Compositions and methods for selectively depleting senescent cells

ABSTRACT

The present disclosure provides compositions and methods for selectively killing senescent cells, wherein the composition comprises piperlongumine (PL) or derivative thereof. The selective killing of senescent cells may delay aging and/or treat age-related disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.15/328,368, filed Jan. 23, 2017, which is the national stage ofInternational application number PCT/US2015/041470, filed Jul. 22, 2015,which claims the benefit of U.S. provisional application No. 62/027,572,filed Jul. 22, 2014, U.S. provisional application No. 62/087,499, filedDec. 4, 2014 and U.S. provisional application No. 62/134,155, filed Mar.17, 2015, each of the disclosures of which are hereby incorporated byreference in its entirety.

GOVERNMENTAL RIGHTS

This invention was made with government support under R01 CA122023awarded by the National Institutes of Health. The government has certainrights in the invention.

FIELD OF THE INVENTION

The present invention relates to piperlongumine and piperlonguminederivatives and their method of use in the treatment and prevention ofdiseases and pathologies related to accumulation of senescent cellsduring aging, such as chronic obstructive pulmonary disease (COPD),osteoarthritis, atherosclerosis, neurodegenerative diseases, diabetes,and many others. The present invention also relates to pharmaceuticalcompositions containing these compounds as well as various uses thereof.

BACKGROUND OF THE INVENTION

Age is a leading risk factor for many human diseases, including mostcancers, atherosclerosis, neurodegenerative diseases, diabetes, and manyothers. An increasing body of evidence demonstrates that aging isassociated with an accumulation of senescent cells. When a cell becomessenescent, it loses its reproductive function, which may cause tissuedegeneration. In addition, senescent cells produce increased levels offree radical and various inflammatory mediators that can induce tissuedamage and cell transformation. Therefore, selective depletion ofsenescent cells may be a novel anti-aging strategy that may preventvarious human diseases associated with aging and rejuvenate the body tolive a healthier lifespan. This assumption is supported by a recentstudy showing that selective depletion of senescent cells in the BubR1progeroid mouse model by a genetic approach resulted in the delay ofvarious age-related pathologies and disorders. However, there is no drugthat can selectively deplete senescent cells. Therefore, a method toselectively deplete senescent cells is needed.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A-E depicts graphs showing piperlongumine (PL) selectively killssenescent WI38 cells but not normal WI38 cells in a dose- andtime-dependent manner. Normal WI38 cells (FIG. 1A), ionizing radiation(IR, 10 Gy) induced senescent WI38 cells (FIG. 1B), or replicativesenescent WI38 cells (FIG. 1C) were incubated with increasingconcentration of PL. Cell viability was measured at 72 h after PLtreatment and expressed as percent of control. Data are presented asmean±SE of three independent assays. IR-induced senescent WI38 cellswere incubated with 10 uM PL and cells viability was measured 1, 3, 6,12, 24, 48 or 72 h after the initiation of PL treatment (FIG. 1D), orthe cells were incubated with 10 uM PL for various durations (1, 3, 6,12, 24, 48 or 72 h) and cell viability was measured at 72 h (FIG. 1E).Data are presented as mean±SE of three independent assays.

FIG. 2A-B depicts graphs showing PL selectively increases reactiveoxygen species (ROS) in senescent WI38 in a time-dependent manner.Normal (FIG. 2A) and IR-induced senescent WI38 cells (FIG. 2B) wereincubated with vehicle (Veh), 200 mM H₂O₂ for 30 min as a positivecontrol, or 10 μM PL. ROS was measured at various times (1, 3, 6, 12, 24hours) after PL treatment and expressed as percent of control. Data arepresented as mean±SE of three independent assays.

FIG. 3A-B depicts graphs showing IR-induced senescent WI38 cells werepre-treated with (FIG. 3A) the antioxidant NAC (1 mM) or (FIG. 3B) thepan-caspase inhibitor Q-VD-OPh (20 μM) for 2 hours and then incubatedwith vehicle (VEH) or 10 μM PL for 72 h. Cell viability was measured andexpressed as percent of control. Data are presented as mean±SE of threeindependent assays.

FIG. 4A-C depicts graphs showing PL and ABT263 can synergistically killsenescent cells but have minimal effect on normal cells. (FIG. 4A)Normal WI38 cells were incubated with vehicle (VEH), 1.25 μM ABT263, PL(10 uM), or both. (FIG. 4B) IR-induced senescent WI38 cells wereincubated with vehicle (VEH), 1.25 μM ABT263, PL (5 or 10 uM), or both.(FIG. 4C) IR-induced senescent WI38 cells were incubated with vehicle(VEH), PL (10 uM), increasing concentrations of ABT263, or both. Cellviability was measured at 72 h and expressed as percent of control. Dataare presented as mean±SE of three independent assays.

FIG. 5A-C depicts graphs showing PL derivative XL-11155B selectivelykills IF-induced senescent WI38 cells but not normal WI38 cells in adose-dependent manner. (FIG. 5A) Structure of XL-11155B. (FIG. 5B)Normal WI38 cells or (FIG. 5C) IR (10 Gy) induced senescent WI38 cellswere incubated with increasing concentrations of XL-11155B. Cellviability was measured at 72 h after XL-11155B treatment and expressedas percent of control. Data are presented as mean±SE of threeindependent assays.

DETAILED DESCRIPTION OF THE INVENTION

Applicants have discovered that piperlongumine (PL), as well asderivatives thereof, selectively kill senescent cells. Accordingly thepresent disclosure provides compositions and methods for selectivelydepleting senescent cells. Additional aspects of the invention aredescribed below.

I. Compositions

In an aspect, a composition of the invention comprises PL or a PLderivative. PL or PL derivatives may be modified to improve potency,bioavailability, solubility, stability, handling properties, or acombination thereof, as compared to an unmodified version. Thus, inanother aspect, a composition of the invention comprises modified PL orPL derivative. In still another aspect, a composition of the inventioncomprises a prodrug of a PL or PL derivative.

A composition of the invention may optionally comprise one or moreadditional drug or therapeutically active agent in addition to the PL orPL derivative. A composition of the invention may further comprise apharmaceutically acceptable excipient, carrier or diluent. Further, acomposition of the invention may contain preserving agents, solubilizingagents, stabilizing agents, wetting agents, emulsifiers, sweeteners,colorants, odorants, salts (substances of the present invention maythemselves be provided in the form of a pharmaceutically acceptablesalt), buffers, coating agents or antioxidants.

Other aspects of the invention are described in further detail below.

(a) Piperlongumine (PL) and PL Derivatives

In general, the compounds detailed herein include compounds comprising aPL, or5,6-dihydro-1-[(2E)-1-oxo-3-(3,4,5-trimethoxyphenyl)-2-propenyl]-2(1H)-pyridinone),structure as diagrammed below. PL may be isolated from a variety ofPiper species (Piperaceae), including Piper aborescens, Pipertuberculatum and the roots of Piper longum L. In addition to extractionof PL from the roots of the Piper plant, PL can also be produced byorganic synthesis (Chatterjee et al., 1967 Tetrahedron 23: 1769-1781).The crystal structure of PL and the adopted conformation of the moleculeare described by Banerjee et al. (Can J. Chem 1986, 64: 867-879).

Provided herein are derivatives of PL. PL derivatives are modifiedversions of PL that are able to selectively deplete senescent cells. Asused herein a “PL derivative” may be a PL derivative known in the art, aPL derivative of Formula (I) or a PL derivative of Formula (II). PLderivatives are known in the art. See for example, US 20090312373, WO2009114126, CN 102125552, CN 102146054, CN 101774875, US 20110053938, WO2012030408, US 20120059004, US 20120157455, US 20130237539, US20140024639, CN 103601670, Nature 475 (2011):231-234, Eur J Med Chem 57(2012):344-361, Tetrahedron 69 (2013):7759-7767, PNAS 109(2012):15115-15120, Eur J Med Chem 82 (2014):545-551, Bioorg Med ChemLett 24 (2014):5727-5730, Journal of Asian Natural Products Research 15(2013):658-669, each of which are incorporated herein in its entirety byreference. PL derivatives with the ability to elevate ROS in senescentcell are potentially used as senolytic drugs.

Provided herein are compounds comprising Formula (I):

wherein:

-   -   X is selected from the group consisting of C(O), C(S), C(NH) and        S(O)₂;    -   Y is selected from the group consisting of O, NH and S;    -   n is an integer from 0-4;    -   R₃, R₄, R₅, R₆, R₇, R₈, R₁₆ and R₁₇ are each independently        selected from the group consisting of hydrogen, deuterium,        halogen, CF₃, CN, OH, OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R,        COR, CO₂R, NOR, NO₂, CONRR, OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R,        NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, a substituted or        unsubstituted C1 to C6 alkyl, a substituted or unsubstituted C1        to C6 alkenyl, a substituted or unsubstituted C1 to C6 alkynyl,        a substituted or unsubstituted aryl, and a substituted or        unsubstituted heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₁₆ and R₁₇, R₁₇ and R₃, R₃ and R₄, R₄ and R₅, R₅        and R₆, R₆ and R₇, or R₇ and R₈ are taken together to form a 4-8        membered saturated or unsaturated ring having 0-3 heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₁₇ and R₃ are eliminated thereby forming a double        bond between the carbons attached to R₄ and R₁₆;    -   A is selected from the group consisting of

wherein:

-   -   R₉ and R₁₀ are independently selected from the group consisting        of hydrogen, deuterium, halogen, CF₃, CN, OH, OCH₃, OR, SR, NRR,        NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR, NO₂, CONRR, OC(O)NRR,        SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, a        substituted or unsubstituted C1 to C6 alkyl, a substituted or        unsubstituted C1 to C6 alkenyl, a substituted or unsubstituted        C1 to C6 alkynyl, a substituted or unsubstituted aryl, and a        substituted or unsubstituted heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₉ and R₁₀ are taken together to form a 5-8 membered        saturated or unsaturated ring having 0-3 heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   B is

wherein:

-   -   R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅ are each independently selected from        the group consisting of hydrogen, deuterium, halogen, CF₃, CN,        OH, OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR,        NO₂, CONRR, OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R,        and C(O)CH₂C(O)R, a substituted or unsubstituted C1 to C6 alkyl,        a substituted or unsubstituted C1 to C6 alkenyl, a substituted        or unsubstituted C1 to C6 alkynyl, a substituted or        unsubstituted aryl, and a substituted or unsubstituted        heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₁₁ and R₁₂, R₁₂ and R₁₃, R₁₃ and R₁₄, and R₁₄ and        R₁₅ are taken together to form a 4-8 membered saturated or        unsaturated ring having 0-3 heteroatoms independently selected        from the group consisting of nitrogen, oxygen, or sulfur;    -   m is an integer from 0-6;    -   Optionally, the phenyl ring in B can be replaced by the        following one or more monocyclic aryl, one or more heteroaryl, a        3-7 membered saturated or partially unsaturated carbocyclic        ring, an 8-10 membered bicyclic saturated, partially unsaturated        or aryl ring, a 5-6 membered monocyclic heteroaryl ring having        1-4 heteroatoms independently selected from nitrogen, oxygen, or        sulfur, a 4-7 membered saturated or partially unsaturated        heterocyclic ring having 1-3 heteroatoms independently selected        from nitrogen, oxygen, or sulfur, a 7-10 membered bicyclic        saturated or partially unsaturated heterocyclic ring having 1-5        heteroatoms independently selected from the group consisting of        nitrogen, oxygen or sulfur, or an 8-10 membered bicyclic        heteroaryl ring having 1-5 heteroatoms independently selected        from the group consisting of nitrogen, oxygen, or sulfur.

In an embodiment, a compound of Formula (I) comprises any of thepreceding compounds of Formula (I), wherein X is selected from the groupconsisting of C(O) and S(O)₂.

In another embodiment, a compound of Formula (I) comprises any of thepreceding compounds of Formula (I), wherein Y is O.

In still another embodiment, a compound of Formula (I) comprises any ofthe preceding compounds of Formula (I), wherein n is an integer from0-2.

In still yet another embodiment, a compound of Formula (I) comprises anyof the preceding compounds of Formula (I), wherein R₃, R₄, R₅, R₆, R₇,R₈, R₁₆ and R₁₇ are each independently selected from the groupconsisting of hydrogen, SR, a substituted or unsubstituted C1 to C6alkyl, a substituted or unsubstituted C1 to C6 alkenyl, a substituted orunsubstituted C1 to C6 alkynyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted heteroaryl, and R₁₇ and R₃ are eliminatedthereby forming a double bond between the carbons attached to R₄ andR₁₆.

In a different embodiment, a compound of Formula (I) comprises any ofthe preceding compounds of Formula (I), wherein A is selected from thegroup consisting of

In other embodiments, a compound of Formula (I) comprises any of thepreceding compounds of Formula (I), wherein R₉ and R₁₀ are independentlyselected from the group consisting of hydrogen, halogen, and CN.

In another embodiment, a compound of Formula (I) comprises any of thepreceding compounds of Formula (I), wherein R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅are each independently selected from the group consisting of hydrogen,halogen, CF₃, OH, OCH₃, OR, NRR, NRCOR, NO₂, and a substituted orunsubstituted C1 to C6 alkyl; and R is independently selected from thegroup consisting of hydrogen, a substituted C1-C4 aliphatic moiety, andan aliphatic moiety containing nitrogen, oxygen, or sulfur; or thephenyl ring in B can be replaced by a 5-6 membered monocyclic heteroarylring having 1-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur, a 7-10 membered bicyclic saturated or partiallyunsaturated heterocyclic ring having 1-5 heteroatoms independentlyselected from the group consisting of nitrogen, oxygen or sulfur, or an8-10 membered bicyclic heteroaryl ring having 1-5 heteroatomsindependently selected from the group consisting of nitrogen, oxygen, orsulfur.

In exemplary embodiments, a compound of the disclosure comprises Formula(II) as depicted in Table 2.

Also provided herein are compounds comprising Formula (II):

wherein:

-   -   X is selected from the group consisting of C(O), C(S), C(NH) and        S(O)₂;    -   Y is selected from the group consisting of O, NH and S;    -   n is an integer from 0-4;    -   R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are each independently        selected from the group consisting of hydrogen, deuterium,        halogen, CF₃, CN, OH, OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R,        COR, CO₂R, NOR, NO₂, CONRR, OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R,        NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, a substituted or        unsubstituted C1 to C6 alkyl, a substituted or unsubstituted C1        to C6 alkenyl, a substituted or unsubstituted C1 to C6 alkynyl,        a substituted or unsubstituted aryl, and a substituted or        unsubstituted heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₁ and R₂, R₂ and R₃, R₃ and R₄, R₄ and R₅, R₅ and        R₆, R₆ and R₇, or R₇ and R₈ are taken together to form a 4-8        membered saturated or unsaturated ring having 0-3 heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   A is selected from the group consisting of

wherein:

-   -   R₉ and R₁₀ are independently selected from the group consisting        of hydrogen, deuterium, halogen, CF₃, CN, OH, OCH₃, OR, SR, NRR,        NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR, NO₂, CONRR, OC(O)NRR,        SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, a        substituted or unsubstituted C1 to C6 alkyl, a substituted or        unsubstituted C1 to C6 alkenyl, a substituted or unsubstituted        C1 to C6 alkynyl, a substituted or unsubstituted aryl, and a        substituted or unsubstituted heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₉ and R₁₀ are taken together to form a 5-8 membered        saturated or unsaturated ring having 0-3 heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   B is

wherein:

-   -   R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅ are each independently selected from        the group consisting of hydrogen, deuterium, halogen, CF₃, CN,        OH, OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR,        NO₂, CONRR, OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R,        and C(O)CH₂C(O)R, a substituted or unsubstituted C1 to C6 alkyl,        a substituted or unsubstituted C1 to C6 alkenyl, a substituted        or unsubstituted C1 to C6 alkynyl, a substituted or        unsubstituted aryl, and a substituted or unsubstituted        heteroaryl;    -   R is independently selected from the group consisting of        hydrogen, substituted C1-C4 aliphatic moiety, aliphatic moiety        containing nitrogen, oxygen, or sulfur, or alternately, two R        moieties bound to the same nitrogen atom are optionally taken        together with the nitrogen atom to form a 3-7 membered saturated        or unsaturated ring having 1-2 additional heteroatoms        independently selected from the group consisting of nitrogen,        oxygen, or sulfur;    -   Optionally, R₁₁ and R₁₂, R₁₂ and R₁₃, R₁₃ and R₁₄, and R₁₄ and        R₁₅ are taken together to form a 4-8 membered saturated or        unsaturated ring having 0-3 heteroatoms independently selected        from the group consisting of nitrogen, oxygen, or sulfur;    -   m is an integer from 0-6;    -   Optionally, the phenyl ring in B can be replaced by the        following one or more monocyclic aryl, one or more heteroaryl, a        3-7 membered saturated or partially unsaturated carbocyclic        ring, an 8-10 membered bicyclic saturated, partially unsaturated        or aryl ring, a 5-6 membered monocyclic heteroaryl ring having        1-4 heteroatoms independently selected from nitrogen, oxygen, or        sulfur, a 4-7 membered saturated or partially unsaturated        heterocyclic ring having 1-3 heteroatoms independently selected        from nitrogen, oxygen, or sulfur, a 7-10 membered bicyclic        saturated or partially unsaturated heterocyclic ring having 1-5        heteroatoms independently selected from the group consisting of        nitrogen, oxygen or sulfur, or an 8-10 membered bicyclic        heteroaryl ring having 1-5 heteroatoms independently selected        from the group consisting of nitrogen, oxygen, or sulfur.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove and R₁ is hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove and R₁ and R₂ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, and R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl.

In yet another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B areas described above and X is selected from the group consisting of C(O)and S(O)₂.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen and X is selected from the groupconsisting of C(O) and S(O)₂.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen and X is selected from the groupconsisting of C(O) and S(O)₂.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl and X is selected from the group consisting of C(O) andS(O)₂.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove and Y is O.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ is hydrogen and Y is O.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen and Y is O.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl and Y is O.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, X is selected from the group consisting of C(O) and S(O)₂ and Yis O.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂ and Y is O.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen, X is selected from the group consistingof C(O) and S(O)₂ and Y is O.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, X is selected from the group consisting of C(O) and S(O)₂and Y is O.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove and n is an integer from 0-2.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen and n is an integer from 0-2.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen and n is an integer from 0-2.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl and n is an integer from 0-2.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, X is selected from the group consisting of C(O) and S(O)₂ and nis an integer from 0-2.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ is hydrogen, X is selected from the group consisting of C(O)and S(O)₂ and n is an integer from 0-2.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ and R₂ are hydrogen, X is selected from the groupconsisting of C(O) and S(O)₂ and n is an integer from 0-2.

In yet still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, X is selected from the group consisting of C(O) and S(O)₂and n is an integer from 0-2.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove and Y is O and n is an integer from 0-2.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, Y is O and n is an integer from 0-2.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen, Y is O and n is an integer from 0-2.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, Y is O and n is an integer from 0-2.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as described above,X is selected from the group consisting of C(O) and S(O)₂, Y is O and nis an integer from 0-2.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, R₃, R₄, R₅, R₆, R₇, R₈, A and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O and n is an integer from 0-2.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, A and B are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O and n is an integer from 0-2.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O and n is aninteger from 0-2.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, A and B are as described above and R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, A and B are as described above, R₁is hydrogen and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, A and B are as described above, R₁ and R₂ arehydrogen and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, and R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, R₂, A and B are as described above, X is selectedfrom the group consisting of C(O) and S(O)₂, and R₃, R₄, R₅, R₆, R₇, R₈are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, A and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, andR₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, A and B are as described above, R₁ and R₂ arehydrogen X, is selected from the group consisting of C(O) and S(O)₂, andR₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, A and B are as described above, Y is O, and R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, A and B are as described above, R₁ ishydrogen, Y is O, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, A and B are as described above, R₁ and R₂ arehydrogen, Y is O, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, and R₃, R₄, R₅, R₆, R₇,R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, A and B are as described above, X is selected fromthe group consisting of C(O) and S(O)₂, Y is O, and R₃, R₄, R₅, R₆, R₇,R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, A and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, Yis O, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, A and B are as describedabove, R₁ and R₂ are hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, and R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, A and B are as described above, n is an integerfrom 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, A and B are as described above, R₁ ishydrogen, n is an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, A and B are as described above, R₁ and R₂ arehydrogen, n is an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, n is an integer from 0-2, andR₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, A and B are as described above, X is selected fromthe group consisting of C(O) and S(O)₂, n is an integer from 0-2, andR₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, A and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, nis an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, A and B are as described above, R₁ and R₂ arehydrogen, X is selected from the group consisting of C(O) and S(O)₂, nis an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, n is an integer from 0-2, and R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, A and B are as described above and Y is O, n is aninteger from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, A and B are as described above, R₁ ishydrogen, Y is O, n is an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, A and B are as described above, R₁ and R₂ arehydrogen, Y is O, n is an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, A and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, n is an integer from0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, A and B are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2,and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, A and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, Yis O, n is an integer from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: A and B are as described above, R₁ and R₂ are hydrogen, Xis selected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, and R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: A and B are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2, and R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, and A is selected from the groupconsisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ and R₂ are hydrogen, and A is selected from the groupconsisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, and Ais selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, and A is selected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ and R₂ are hydrogen, X is selected from the group consistingof C(O) and S(O)₂, and A is selected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, X is selected from the group consisting of C(O) and S(O)₂,and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, Y is O, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, Y is O, and A is selected from thegroup consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ and R₂ are hydrogen, Y is O, and A is selected from the groupconsisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, Y is O, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, Y isO, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, and A is selected from the group consistingof

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O, and A is selected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, and A isselected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, n is an integer from 0-2, and A is selected from the groupconsisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, n is an integer from 0-2, and A isselected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ and R₂ are hydrogen, n is an integer from 0-2, and A isselected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, n is an integer from 0-2, and A is selected from the groupconsisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, n isan integer from 0-2, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, n is an integer from 0-2, and A is selected from thegroup consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, n is an integer from 0-2, and A is selected from the groupconsisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, n is an integerfrom 0-2, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described aboveand Y is O, n is an integer from 0-2, and A is selected from the groupconsisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, Y is O, n is an integer from 0-2, and Ais selected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above,R₁ and R₂ are hydrogen, Y is O, n is an integer from 0-2, and A isselected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, Y isO, n is an integer from 0-2, and A is selected from the group consistingof

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above, Xis selected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, R₃, R₄, R₅, R₆, R₇, R₈, and B are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, n is an integer from 0-2, and A is selectedfrom the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above, R₁and R₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, Y is O, n is an integer from 0-2, and A is selected from thegroup consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, and B are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, and B are as described above, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, and B are as described above, R₁ ishydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected fromthe group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, and B are as described above, R₁ and R₂ arehydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected fromthe group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, R₂, and B are as described above, X is selected fromthe group consisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from the groupconsisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, and B are as described above, R₁ and R₂ arehydrogen X, is selected from the group consisting of C(O) and S(O)₂, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from the groupconsisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and Ais selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, and B are as described above, Y is O, R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, and B are as described above, R₁ ishydrogen, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selectedfrom the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, and B are as described above, R₁ and R₂ arehydrogen, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selectedfrom the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, R₃, R₄, R₅, R₆, R₇, R₈are hydrogen, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, and B are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and A is selected from the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, Yis O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from thegroup consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and B are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A isselected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, and B are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, and B are as described above, n is an integerfrom 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected fromthe group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, and B are as described above, R₁ ishydrogen, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and A is selected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, and B are as described above, R₁ and R₂ arehydrogen, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and A is selected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, and B are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, n is an integer from 0-2, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from the groupconsisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, and B are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, n is an integer from 0-2, R₃, R₄,R₅, R₆, R₇, R₈ are hydrogen, and A is selected from the group consistingof

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, and B are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, nis an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A isselected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, and B are as described above, R₁ and R₂ are hydrogen,X is selected from the group consisting of C(O) and S(O)₂, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selectedfrom the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, and B are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, n is an integer from 0-2, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and A is selected from the group consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, and B are as described above and Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selectedfrom the group consisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, and B are as described above, R₁ ishydrogen, Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and A is selected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, and B are as described above, R₁ and R₂ are hydrogen,Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and A is selected from the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, and B are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, n is an integer from0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from thegroup consisting of

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, and B are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2,R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selected from the groupconsisting of

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, and B are as described above, R₁ is hydrogen,X is selected from the group consisting of C(O) and S(O)₂, Y is O, n isan integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A isselected from the group consisting of

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: B is as described above, R₁ and R₂ are hydrogen, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and A is selectedfrom the group consisting of

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: B is as described above, R₁ is hydrogen, R₂ isselected from the group consisting of a substituted or unsubstituted C1to C6 alkyl, a substituted or unsubstituted aryl, and a substituted orunsubstituted heteroaryl, X is selected from the group consisting ofC(O) and S(O)₂, Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈are hydrogen, and A is selected from the group consisting of

Any of the preceding compounds wherein R₉ and R₁₀ are hydrogen.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, and B is as described above wherein:R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ and R₂ are hydrogen, and B is as described above wherein: R₁₁and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, and Bis as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ andR₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ and R₂ are hydrogen, X is selected from the group consistingof C(O) and S(O)₂, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, X is selected from the group consisting of C(O) and S(O)₂,and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, Y is O, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, Y is O, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ and R₂ are hydrogen, Y is O, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, Y is O, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, Y isO, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂,R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, and B is as described above wherein: R₁₁ andR₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, and B isas described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, n is an integer from 0-2, and B is as described above wherein:R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, n is an integer from 0-2, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ and R₂ are hydrogen, n is an integer from 0-2, B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, n is an integer from 0-2, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, n isan integer from 0-2, and B is as described above wherein: R₁₁ and R₁₅are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, n is an integer from 0-2, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, n is an integer from 0-2, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, n is an integerfrom 0-2, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described aboveand Y is O, n is an integer from 0-2, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, Y is O, n is an integer from 0-2, and Bis as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ andR₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above,R₁ and R₂ are hydrogen, Y is O, n is an integer from 0-2, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, Y isO, n is an integer from 0-2, and B is as described above wherein: R₁₁and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above, Xis selected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, R₃, R₄, R₅, R₆, R₇, R₈, and A are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, n is an integer from 0-2, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above, R₁and R₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, Y is O, n is an integer from 0-2, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₃, R₄, R₅, R₆, R₇, R₈, and A are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, and A are as described above, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁ and R₁₅are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, and A are as described above, R₁ ishydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as describedabove wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and Ris as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, and A are as described above, R₁ and R₂ arehydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as describedabove wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and Ris as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, and A are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₁, R₂, and A are as described above, X isselected from the group consisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁ and R₁₅are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₂, and A are as described above, R₁ is hydrogen, X isselected from the group consisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁ and R₁₅are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, and A are as described above, R₁ and R₂ arehydrogen X, is selected from the group consisting of C(O) and S(O)₂, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as described above wherein:R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, and A are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and Bis as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ andR₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, and A are as described above, Y is O, R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁ andR₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, and A are as described above, R₁ ishydrogen, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, and A are as described above, R₁ and R₂ arehydrogen, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, and A are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, R₃, R₄, R₅, R₆, R₇, R₈are hydrogen, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, and A are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, and A are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, Yis O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, R₈, and A are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, and A are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, and A are as described above, n is an integerfrom 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as describedabove wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and Ris as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, and A are as described above, R₁ ishydrogen, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, and A are as described above, R₁ and R₂ arehydrogen, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, and A are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, n is an integer from 0-2, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as described above wherein:R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, and A are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, n is an integer from 0-2, R₃, R₄,R₅, R₆, R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as describedabove.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, and A are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, nis an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, and A are as described above, R₁ and R₂ are hydrogen,X is selected from the group consisting of C(O) and S(O)₂, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, and A are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, n is an integer from 0-2, R₃, R₄, R₅, R₆,R₇, R₈ are hydrogen, and B is as described above wherein: R₁₁ and R₁₅are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, and A are as described above and Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, and A are as described above, R₁ ishydrogen, Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, and B is as described above wherein: R₁₁ and R₁₅ are hydrogen;R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, and A are as described above, R₁ and R₂ are hydrogen,Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, and A are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, n is an integer from0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, and A are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2,R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is as described abovewherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is asdescribed above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, and A are as described above, R₁ is hydrogen,X is selected from the group consisting of C(O) and S(O)₂, Y is O, n isan integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: A is as described above, R₁ and R₂ are hydrogen, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, and B is asdescribed above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃ and R₁₄ areOR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: A is as described above, R₁ is hydrogen, R₂ isselected from the group consisting of a substituted or unsubstituted C1to C6 alkyl, a substituted or unsubstituted aryl, and a substituted orunsubstituted heteroaryl, X is selected from the group consisting ofC(O) and S(O)₂, Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈are hydrogen, and B is as described above wherein: R₁₁ and R₁₅ arehydrogen; R₁₂, R₁₃ and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ and R₂ are hydrogen, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, X is selected from the group consisting of C(O) and S(O)₂, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above,R₁ and R₂ are hydrogen, X is selected from the group consisting of C(O)and S(O)₂, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, A is selected fromthe group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, Y is O, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, Y is O, A is selected from the groupconsisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above,R₁ and R₂ are hydrogen, Y is O, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, Y isO, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, Xis selected from the group consisting of C(O) and S(O)₂, Y is O, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, Y is O, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, R₁and R₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, Y is O, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, n is an integer from 0-2, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, n is an integer from 0-2, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above,R₁ and R₂ are hydrogen, n is an integer from 0-2, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, n isan integer from 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, Xis selected from the group consisting of C(O) and S(O)₂, n is an integerfrom 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, X is selected from the group consistingof C(O) and S(O)₂, n is an integer from 0-2, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, R₁and R₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, n is an integer from 0-2, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, n is an integerfrom 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as described aboveand Y is O, n is an integer from 0-2, A is selected from the groupconsisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are asdescribed above, R₁ is hydrogen, Y is O, n is an integer from 0-2, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, R₁and R₂ are hydrogen, Y is O, n is an integer from 0-2, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, Y isO, n is an integer from 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, X is selected from the group consisting of C(O)and S(O)₂, Y is O, n is an integer from 0-2, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, R₁ andR₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, Y is O, n is an integer from 0-2, A is selected from the groupconsisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₃, R₄, R₅, R₆, R₇, and R₈ are as describedabove, R₁ is hydrogen, R₂ is selected from the group consisting of asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted aryl, and a substituted or unsubstituted heteroaryl, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, n, R₁, and R₂ are as described above, R₃, R₄, R₅, R₆, R₇,R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, n, and R₂ are as described above, R₁ ishydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, Y, and n are as described above, R₁ and R₂ arehydrogen, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, and n are as described above, R₁ ishydrogen, R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, n, R₁, and R₂ are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, n, and R₂ are as described above, R₁ ishydrogen, X is selected from the group consisting of C(O) and S(O)₂, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y, and n are as described above, R₁ and R₂ are hydrogenX, is selected from the group consisting of C(O) and S(O)₂, R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, and n are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, X is selected from the groupconsisting of C(O) and S(O)₂, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, n, R₁, and R₂ are as described above, Y is O, R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, n, and R₂ are as described above, R₁ ishydrogen, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, and n are as described above, R₁ and R₂ are hydrogen,Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, and n are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, Y is O, R₃, R₄, R₅, R₆, R₇, R₈are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: n, R₁, and R₂ are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: n, and R₂ are as described above, R₁ is hydrogen,X is selected from the group consisting of C(O) and S(O)₂, Y is O, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: n, R₃, R₄, R₅, R₆, R₇, and R₈ are as described above, R₁and R₂ are hydrogen, X is selected from the group consisting of C(O) andS(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected fromthe group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: n is as described above, R₁ is hydrogen, R₂ isselected from the group consisting of a substituted or unsubstituted C1to C6 alkyl, a substituted or unsubstituted aryl, and a substituted orunsubstituted heteroaryl, X is selected from the group consisting ofC(O) and S(O)₂, Y is O, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, Y, R₁, and R₂ are as described above, n is an integer from0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from the groupconsisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, Y, and R₂ are as described above, R₁ ishydrogen, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen,A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X, and Y are as described above, R₁ and R₂ are hydrogen,n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, and Y are as described above, R₁ is hydrogen,R₂ is selected from the group consisting of a substituted orunsubstituted C1 to C6 alkyl, a substituted or unsubstituted aryl, and asubstituted or unsubstituted heteroaryl, n is an integer from 0-2, R₃,R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consistingof

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: Y, R₁, and R₂ are as described above, X is selected from thegroup consisting of C(O) and S(O)₂, n is an integer from 0-2, R₃, R₄,R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y, and R₂ are as described above, R₁ is hydrogen,X is selected from the group consisting of C(O) and S(O)₂, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₆ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: Y is as described above, R₁ and R₂ are hydrogen, X isselected from the group consisting of C(O) and S(O)₂, n is an integerfrom 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selected from thegroup consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: Y is as described above, R₁ is hydrogen, R₂ isselected from the group consisting of a substituted or unsubstituted C1to C6 alkyl, a substituted or unsubstituted aryl, and a substituted orunsubstituted heteroaryl, X is selected from the group consisting ofC(O) and S(O)₂, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ arehydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: X, R₁, and R₂ are as described above and Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: X, and R₂ are as described above, R₁ is hydrogen,Y is O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, Ais selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: X is as described above, R₁ and R₂ are hydrogen, Y is O,n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A isselected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: X is as described above, R₁ is hydrogen, R₂ isselected from the group consisting of a substituted or unsubstituted C1to C6 alkyl, a substituted or unsubstituted aryl, and a substituted orunsubstituted heteroaryl, Y is O, n is an integer from 0-2, R₃, R₄, R₅,R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In one embodiment, a compound of the disclosure comprises Formula (II),wherein: R₁, and R₂ are as described above, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2, R₃, R₄,R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In a different embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₂ is as described above, R₁ is hydrogen, X isselected from the group consisting of C(O) and S(O)₂, Y is O, n is aninteger from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, A is selectedfrom the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In another embodiment, a compound of the disclosure comprises Formula(II), wherein: R₁ and R₂ are hydrogen, X is selected from the groupconsisting of C(O) and S(O)₂, Y is O, n is an integer from 0-2, R₃, R₄,R₅, R₆, R₇, R₈ are hydrogen, A is selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

In still another embodiment, a compound of the disclosure comprisesFormula (II), wherein: R₁ is hydrogen, R₂ is selected from the groupconsisting of a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted aryl, and a substituted or unsubstitutedheteroaryl, X is selected from the group consisting of C(O) and S(O)₂, Yis O, n is an integer from 0-2, R₃, R₄, R₅, R₆, R₇, R₈ are hydrogen, Ais selected from the group consisting of

and B is as described above wherein: R₁₁ and R₁₅ are hydrogen; R₁₂, R₁₃and R₁₄ are OR; and R is as described above.

Any of the preceding compounds wherein R₉ and R₁₀ are hydrogen.

Any of the preceding compounds wherein R₁₂, R₁₃ and R₁₄ are OCH₃.

In exemplary embodiments, a compound of the disclosure comprises Formula(II) as shown in Table 2.

Dosages of a PL, a PL derivative, a compound of Formula (I) or acompound of Formula (II) can vary between wide limits, depending uponthe disease or disorder to be treated, the age and condition of thesubject to be treated. In an embodiment where a composition comprising aPL, a PL derivative, a compound of Formula (I) or a compound of Formula(II) is contacted with a sample, the concentration of the PL, a PLderivative, a compound of Formula (I) or a compound of Formula (II) maybe from about 1 μM to about 40 μM. Alternatively, the concentration ofthe PL, a PL derivative, a compound of Formula (I) or a compound ofFormula (II) may be from about 5 μM to about 25 μM. For example, theconcentration of the PL, a PL derivative, a compound of Formula (I) or acompound of Formula (II) may be about 1, about 2.5 about 5, about 6,about 7, about 8, about 9, about 10, about 11, about 12, about 12, about14, about 15, about 16, about 17, about 18, about 19, about 20, about21, about 22, about 23, about 24, about 25, about 30, about 35, or about40 μM. Additionally, the concentration of the PL, a PL derivative, acompound of Formula (I) or a compound of Formula (II) may be greaterthan 40 μM. For example, the concentration of the PL, a PL derivative, acompound of Formula (I) or a compound of Formula (II) may be about 40,about 45, about 50, about 55, about 60, about 65, about 70, about 75,about 80, about 85, about 90, about 95 or about 100 μM.

In an embodiment where the composition comprising a PL, a PL derivative,a compound of Formula (I) or a compound of Formula (II) is administeredto a subject, the dose of the PL, a PL derivative, a compound of Formula(I) or a compound of Formula (II) may be from about 0.1 mg/kg to about500 mg/kg. For example, the dose of a PL, a PL derivative, a compound ofFormula (I) or a compound of Formula (II) may be about 0.1 mg/kg, about0.5 mg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg,about 20 mg/kg, or about 25 mg/kg. Alternatively, the dose of the PL, aPL derivative, a compound of Formula (I) or a compound of Formula (II)may be about 25 mg/kg, about 50 mg/kg, about 75 mg/kg, about 100 mg/kg,about 125 mg/kg, about 150 mg/kg, about 175 mg/kg, about 200 mg/kg,about 225 mg/kg, or about 250 mg/kg. Additionally, the dose of the PL, aPL derivative, a compound of Formula (I) or a compound of Formula (II)may be about 300 mg/kg, about 325 mg/kg, about 350 mg/kg, about 375mg/kg, about 400 mg/kg, about 425 mg/kg, about 450 mg/kg, about 475mg/kg or about 500 mg/kg.

(b) Components of the Composition

The present disclosure also provides pharmaceutical compositions. Thepharmaceutical composition comprises PL, a PL derivative, a compound ofFormula (I) or a compound of Formula (II), as an active ingredient, andat least one pharmaceutically acceptable excipient.

The pharmaceutically acceptable excipient may be a diluent, a binder, afiller, a buffering agent, a pH modifying agent, a disintegrant, adispersant, a preservative, a lubricant, taste-masking agent, aflavoring agent, or a coloring agent. The amount and types of excipientsutilized to form pharmaceutical compositions may be selected accordingto known principles of pharmaceutical science.

In one embodiment, the excipient may be a diluent. The diluent may becompressible (i.e., plastically deformable) or abrasively brittle.Non-limiting examples of suitable compressible diluents includemicrocrystalline cellulose (MCC), cellulose derivatives, cellulosepowder, cellulose esters (i.e., acetate and butyrate mixed esters),ethyl cellulose, methyl cellulose, hydroxypropyl cellulose,hydroxypropyl methylcellulose, sodium carboxymethylcellulose, cornstarch, phosphated corn starch, pregelatinized corn starch, rice starch,potato starch, tapioca starch, starch-lactose, starch-calcium carbonate,sodium starch glycolate, glucose, fructose, lactose, lactosemonohydrate, sucrose, xylose, lactitol, mannitol, malitol, sorbitol,xylitol, maltodextrin, and trehalose. Non-limiting examples of suitableabrasively brittle diluents include dibasic calcium phosphate (anhydrousor dihydrate), calcium phosphate tribasic, calcium carbonate, andmagnesium carbonate.

In another embodiment, the excipient may be a binder. Suitable bindersinclude, but are not limited to, starches, pregelatinized starches,gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodiumcarboxymethylcellulose, ethylcellulose, polyacrylamides,polyvinyloxoazolidone, polyvinylalcohols, C₁₂-C₁₈ fatty acid alcohol,polyethylene glycol, polyols, saccharides, oligosaccharides,polypeptides, oligopeptides, and combinations thereof.

In another embodiment, the excipient may be a filler. Suitable fillersinclude, but are not limited to, carbohydrates, inorganic compounds, andpolyvinylpyrrolidone. By way of non-limiting example, the filler may becalcium sulfate, both di- and tri-basic, starch, calcium carbonate,magnesium carbonate, microcrystalline cellulose, dibasic calciumphosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc,modified starches, lactose, sucrose, mannitol, or sorbitol.

In still another embodiment, the excipient may be a buffering agent.Representative examples of suitable buffering agents include, but arenot limited to, phosphates, carbonates, citrates, tris buffers, andbuffered saline salts (e.g., Tris buffered saline or phosphate bufferedsaline).

In various embodiments, the excipient may be a pH modifier. By way ofnon-limiting example, the pH modifying agent may be sodium carbonate,sodium bicarbonate, sodium citrate, citric acid, or phosphoric acid.

In a further embodiment, the excipient may be a disintegrant. Thedisintegrant may be non-effervescent or effervescent. Suitable examplesof non-effervescent disintegrants include, but are not limited to,starches such as corn starch, potato starch, pregelatinized and modifiedstarches thereof, sweeteners, clays, such as bentonite,micro-crystalline cellulose, alginates, sodium starch glycolate, gumssuch as agar, guar, locust bean, karaya, pecitin, and tragacanth.Non-limiting examples of suitable effervescent disintegrants includesodium bicarbonate in combination with citric acid and sodiumbicarbonate in combination with tartaric acid.

In yet another embodiment, the excipient may be a dispersant ordispersing enhancing agent. Suitable dispersants may include, but arenot limited to, starch, alginic acid, polyvinylpyrrolidones, guar gum,kaolin, bentonite, purified wood cellulose, sodium starch glycolate,isoamorphous silicate, and microcrystalline cellulose.

In another alternate embodiment, the excipient may be a preservative.Non-limiting examples of suitable preservatives include antioxidants,such as BHA, BHT, vitamin A, vitamin C, vitamin E, or retinyl palmitate,citric acid, sodium citrate; chelators such as EDTA or EGTA; andantimicrobials, such as parabens, chlorobutanol, or phenol.

In a further embodiment, the excipient may be a lubricant. Non-limitingexamples of suitable lubricants include minerals such as talc or silica;and fats such as vegetable stearin, magnesium stearate or stearic acid.

In yet another embodiment, the excipient may be a taste-masking agent.Taste-masking materials include cellulose ethers; polyethylene glycols;polyvinyl alcohol; polyvinyl alcohol and polyethylene glycol copolymers;monoglycerides or triglycerides; acrylic polymers; mixtures of acrylicpolymers with cellulose ethers; cellulose acetate phthalate; andcombinations thereof.

In an alternate embodiment, the excipient may be a flavoring agent.Flavoring agents may be chosen from synthetic flavor oils and flavoringaromatics and/or natural oils, extracts from plants, leaves, flowers,fruits, and combinations thereof.

In still a further embodiment, the excipient may be a coloring agent.Suitable color additives include, but are not limited to, food, drug andcosmetic colors (FD&C), drug and cosmetic colors (D&C), or external drugand cosmetic colors (Ext. D&C).

The weight fraction of the excipient or combination of excipients in thecomposition may be about 99% or less, about 97% or less, about 95% orless, about 90% or less, about 85% or less, about 80% or less, about 75%or less, about 70% or less, about 65% or less, about 60% or less, about55% or less, about 50% or less, about 45% or less, about 40% or less,about 35% or less, about 30% or less, about 25% or less, about 20% orless, about 15% or less, about 10% or less, about 5% or less, about 2%,or about 1% or less of the total weight of the composition.

The composition can be formulated into various dosage forms andadministered by a number of different means that will deliver atherapeutically effective amount of the active ingredient. Suchcompositions can be administered orally, parenterally, or topically indosage unit formulations containing conventional nontoxicpharmaceutically acceptable carriers, adjuvants, and vehicles asdesired. Topical administration may also involve the use of transdermaladministration such as transdermal patches or iontophoresis devices. Theterm parenteral as used herein includes subcutaneous, intravenous,intramuscular, or intrasternal injection, or infusion techniques.Formulation of drugs is discussed in, for example, Gennaro, A. R.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.(18^(th) ed, 1995), and Liberman, H. A. and Lachman, L., Eds.,Pharmaceutical Dosage Forms, Marcel Dekker Inc., New York, N.Y. (1980).In a specific embodiment, a composition may be a food supplement or acomposition may be a cosmetic.

Solid dosage forms for oral administration include capsules, tablets,caplets, pills, powders, pellets, and granules. In such solid dosageforms, the active ingredient is ordinarily combined with one or morepharmaceutically acceptable excipients, examples of which are detailedabove. Oral preparations may also be administered as aqueoussuspensions, elixirs, or syrups. For these, the active ingredient may becombined with various sweetening or flavoring agents, coloring agents,and, if so desired, emulsifying and/or suspending agents, as well asdiluents such as water, ethanol, glycerin, and combinations thereof.

For parenteral administration (including subcutaneous, intradermal,intravenous, intramuscular, and intraperitoneal), the preparation may bean aqueous or an oil-based solution. Aqueous solutions may include asterile diluent such as water, saline solution, a pharmaceuticallyacceptable polyol such as glycerol, propylene glycol, or other syntheticsolvents; an antibacterial and/or antifungal agent such as benzylalcohol, methyl paraben, chlorobutanol, phenol, thimerosal, and thelike; an antioxidant such as ascorbic acid or sodium bisulfite; achelating agent such as etheylenediaminetetraacetic acid; a buffer suchas acetate, citrate, or phosphate; and/or an agent for the adjustment oftonicity such as sodium chloride, dextrose, or a polyalcohol such asmannitol or sorbitol. The pH of the aqueous solution may be adjustedwith acids or bases such as hydrochloric acid or sodium hydroxide.Oil-based solutions or suspensions may further comprise sesame, peanut,olive oil, or mineral oil. The compositions may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carried, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets.

For topical (e.g., transdermal or transmucosal) administration,penetrants appropriate to the barrier to be permeated are generallyincluded in the preparation. Pharmaceutical compositions adapted fortopical administration may be formulated as ointments, creams,suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosolsor oils. In some embodiments, the pharmaceutical composition is appliedas a topical ointment or cream. When formulated in an ointment, theactive ingredient may be employed with either a paraffinic or awater-miscible ointment base. Alternatively, the active ingredient maybe formulated in a cream with an oil-in-water cream base or awater-in-oil base. Pharmaceutical compositions adapted for topicaladministration to the eye include eye drops wherein the activeingredient is dissolved or suspended in a suitable carrier, especiallyan aqueous solvent. Pharmaceutical compositions adapted for topicaladministration in the mouth include lozenges, pastilles and mouthwashes. Transmucosal administration may be accomplished through the useof nasal sprays, aerosol sprays, tablets, or suppositories, andtransdermal administration may be via ointments, salves, gels, patches,or creams as generally known in the art.

In certain embodiments, a composition comprising PL, a PL derivative, acompound of Formula (I) or a compound of Formula (II) is encapsulated ina suitable vehicle to either aid in the delivery of the compound totarget cells, to increase the stability of the composition, or tominimize potential toxicity of the composition. As will be appreciatedby a skilled artisan, a variety of vehicles are suitable for deliveringa composition of the present invention. Non-limiting examples ofsuitable structured fluid delivery systems may include nanoparticles,liposomes, microemulsions, micelles, dendrimers and otherphospholipid-containing systems. Methods of incorporating compositionsinto delivery vehicles are known in the art.

In one alternative embodiment, a liposome delivery vehicle may beutilized. Liposomes, depending upon the embodiment, are suitable fordelivery of the PL, a PL derivative, a compound of Formula (I) or acompound of Formula (II) in view of their structural and chemicalproperties. Generally speaking, liposomes are spherical vesicles with aphospholipid bilayer membrane. The lipid bilayer of a liposome may fusewith other bilayers (e.g., the cell membrane), thus delivering thecontents of the liposome to cells. In this manner, the PL or compoundcomprising Formula (I) may be selectively delivered to a cell byencapsulation in a liposome that fuses with the targeted cell'smembrane.

Liposomes may be comprised of a variety of different types ofphosolipids having varying hydrocarbon chain lengths. Phospholipidsgenerally comprise two fatty acids linked through glycerol phosphate toone of a variety of polar groups. Suitable phospholids includephosphatidic acid (PA), phosphatidylserine (PS), phosphatidylinositol(PI), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG),phosphatidylcholine (PC), and phosphatidylethanolamine (PE). The fattyacid chains comprising the phospholipids may range from about 6 to about26 carbon atoms in length, and the lipid chains may be saturated orunsaturated. Suitable fatty acid chains include (common name presentedin parentheses) n-dodecanoate (laurate), n-tretradecanoate (myristate),n-hexadecanoate (palmitate), n-octadecanoate (stearate), n-eicosanoate(arachidate), n-docosanoate (behenate), n-tetracosanoate (lignocerate),cis-9-hexadecenoate (palmitoleate), cis-9-octadecanoate (oleate),cis,cis-9,12-octadecandienoate (linoleate), allcis-9,12,15-octadecatrienoate (linolenate), and allcis-5,8,11,14-eicosatetraenoate (arachidonate). The two fatty acidchains of a phospholipid may be identical or different. Acceptablephospholipids include dioleoyl PS, dioleoyl PC, distearoyl PS,distearoyl PC, dimyristoyl PS, dimyristoyl PC, dipalmitoyl PG, stearoyl,oleoyl PS, palmitoyl, linolenyl PS, and the like.

The phospholipids may come from any natural source, and, as such, maycomprise a mixture of phospholipids. For example, egg yolk is rich inPC, PG, and PE, soy beans contains PC, PE, PI, and PA, and animal brainor spinal cord is enriched in PS. Phospholipids may come from syntheticsources too. Mixtures of phospholipids having a varied ratio ofindividual phospholipids may be used. Mixtures of differentphospholipids may result in liposome compositions having advantageousactivity or stability of activity properties. The above mentionedphospholipids may be mixed, in optimal ratios with cationic lipids, suchas N-(1-(2,3-dioleolyoxy)propyl)-N,N,N-trimethyl ammonium chloride,1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchloarate,3,3′-deheptyloxacarbocyanine iodide,1,1′-dedodecyl-3,3,3′,3′-tetramethylindocarbocyanine perchloarate,1,1′-dioleyl-3,3,3′,3′-tetramethylindo carbocyanine methanesulfonate,N-4-(delinoleylaminostyryl)-N-methylpyridinium iodide, or1,1,-dilinoleyl-3,3,3′,3′-tetramethylindocarbocyanine perchloarate.

Liposomes may optionally comprise sphingolipids, in which spingosine isthe structural counterpart of glycerol and one of the one fatty acids ofa phosphoglyceride, or cholesterol, a major component of animal cellmembranes. Liposomes may optionally contain pegylated lipids, which arelipids covalently linked to polymers of polyethylene glycol (PEG). PEGsmay range in size from about 500 to about 10,000 daltons.

Liposomes may further comprise a suitable solvent. The solvent may be anorganic solvent or an inorganic solvent. Suitable solvents include, butare not limited to, dimethylsulfoxide (DMSO), methylpyrrolidone,N-methylpyrrolidone, acetronitrile, alcohols, dimethylformamide,tetrahydrofuran, or combinations thereof.

Liposomes carrying the PL, PL derivative, compound of Formula (I) orcompound of Formula (II) (i.e., having at least one methionine compound)may be prepared by any known method of preparing liposomes for drugdelivery, such as, for example, detailed in U.S. Pat. Nos. 4,241,046,4,394,448, 4,529,561, 4,755,388, 4,828,837, 4,925,661, 4,954,345,4,957,735, 5,043,164, 5,064,655, 5,077,211 and 5,264,618, thedisclosures of which are hereby incorporated by reference in theirentirety. For example, liposomes may be prepared by sonicating lipids inan aqueous solution, solvent injection, lipid hydration, reverseevaporation, or freeze drying by repeated freezing and thawing. In apreferred embodiment the liposomes are formed by sonication. Theliposomes may be multilamellar, which have many layers like an onion, orunilamellar. The liposomes may be large or small. Continued high-shearsonication tends to form smaller unilamellar lipsomes.

As would be apparent to one of ordinary skill, all of the parametersthat govern liposome formation may be varied. These parameters include,but are not limited to, temperature, pH, concentration of methioninecompound, concentration and composition of lipid, concentration ofmultivalent cations, rate of mixing, presence of and concentration ofsolvent.

In another embodiment, a composition of the invention may be deliveredto a cell as a microemulsion. Microemulsions are generally clear,thermodynamically stable solutions comprising an aqueous solution, asurfactant, and “oil.” The “oil” in this case, is the supercriticalfluid phase. The surfactant rests at the oil-water interface. Any of avariety of surfactants are suitable for use in microemulsionformulations including those described herein or otherwise known in theart. The aqueous microdomains suitable for use in the inventiongenerally will have characteristic structural dimensions from about 5 nmto about 100 nm. Aggregates of this size are poor scatterers of visiblelight and hence, these solutions are optically clear. As will beappreciated by a skilled artisan, microemulsions can and will have amultitude of different microscopic structures including sphere, rod, ordisc shaped aggregates. In one embodiment, the structure may bemicelles, which are the simplest microemulsion structures that aregenerally spherical or cylindrical objects. Micelles are like drops ofoil in water, and reverse micelles are like drops of water in oil. In analternative embodiment, the microemulsion structure is the lamellae. Itcomprises consecutive layers of water and oil separated by layers ofsurfactant. The “oil” of microemulsions optimally comprisesphospholipids. Any of the phospholipids detailed above for liposomes aresuitable for embodiments directed to microemulsions. The PL or compoundcomprising Formula (I) may be encapsulated in a microemulsion by anymethod generally known in the art.

In yet another embodiment, a PL, a PL derivative, a compound of Formula(I) or a compound of Formula (II) may be delivered in a dendriticmacromolecule, or a dendrimer. Generally speaking, a dendrimer is abranched tree-like molecule, in which each branch is an interlinkedchain of molecules that divides into two new branches (molecules) aftera certain length. This branching continues until the branches(molecules) become so densely packed that the canopy forms a globe.Generally, the properties of dendrimers are determined by the functionalgroups at their surface. For example, hydrophilic end groups, such ascarboxyl groups, would typically make a water-soluble dendrimer.Alternatively, phospholipids may be incorporated in the surface of adendrimer to facilitate absorption across the skin. Any of thephospholipids detailed for use in liposome embodiments are suitable foruse in dendrimer embodiments. Any method generally known in the art maybe utilized to make dendrimers and to encapsulate compositions of theinvention therein. For example, dendrimers may be produced by aniterative sequence of reaction steps, in which each additional iterationleads to a higher order dendrimer. Consequently, they have a regular,highly branched 3D structure, with nearly uniform size and shape.Furthermore, the final size of a dendrimer is typically controlled bythe number of iterative steps used during synthesis. A variety ofdendrimer sizes are suitable for use in the invention. Generally, thesize of dendrimers may range from about 1 nm to about 100 nm.

(c) Bcl-2 Family Inhibitor

In an aspect, the composition further comprises at least one inhibitorof one or more anti-apoptotic proteins in the Bcl-2 family. As usedherein, a “Bcl-2 inhibitor” includes at least one inhibitor of one ormore anti-apoptotic proteins in the Bcl-2 family. Specifically, a Bcl-2inhibitor of the invention selectively kills senescent cells. Methods todetermine if a compound inhibits one or more anti-apoptotic proteins inthe Bcl-2 family are known in the art. For example, nucleic acidexpression, protein expression, or activity of Bcl-2 family proteins maybe measured as described in detail below. Methods to determine if acompound selectively kills senescent cells are known in the art. Forexample, see Section III(b) and Section III(c).

Members of the B-cell lymphoma 2 (Bcl-2) family control the integrity ofthe outer mitochondrial membrane (OMM) and thus are critical indetermining the susceptibility of cells to apoptosis induced by theintrinsic pathway. Bcl-2 family members can be divided into threesubfamilies based on structural and functional features: ananti-apoptotic family, a multidomain pro-apoptotic family, and aBH3-only pro-apoptotic family. The anti-apoptotic subfamily suppressesapoptosis and promotes cell survival but not cell proliferation. Assuch, the anti-apoptotic proteins in the Bcl-2 family may also bereferred to as pro-survival proteins. Non-limiting examples ofanti-apoptotic Bcl-2 family proteins may include Bcl-2, Bcl-xl, Bcl-w,Mcl-1, Bfl1/A-1, and Bcl-B. The anti-apoptotic Bcl-2 family proteins arecharacterized by the presence of up to four relatively short sequencemotifs, which are less than 20 amino acids in length, known as Bcl-2homology 1 (BH1), BH2, BH3 and BH4 domains. They also have a C-terminalmembrane-anchoring sequence and a similar three-dimensional structure.Inhibitors of one or more anti-apoptotic proteins in the Bcl-2 familymay promote cell death by antagonizing the pro-survival function of theBcl-2 protein family thereby inducing apoptosis. An inhibitor of theinvention may inhibit one or more anti-apoptotic proteins in the Bcl-2family. Accordingly, an inhibitor of the invention may inhibit one ormore anti-apoptotic proteins selected from the group consisting ofBcl-2, Bcl-xl, Bcl-w, Mcl-1, Bfl1/A-1, and Bcl-B. In certainembodiments, an inhibitor of the invention is a Bcl-2, Bcl-xl and Bcl-winhibitor. In a specific embodiment, an inhibitor of the invention is aBcl-2 and Bcl-xl inhibitor. It is understood that an inhibitor of theinvention may primarily inhibit Bcl-2 and/or Bcl-xl, but also haveinhibitory effects on other members of the anti-apoptotic proteins inthe Bcl-2 family.

An inhibitor of one or more anti-apoptotic proteins in the Bcl-2 familymay be an inhibitor that inhibits nucleic acid expression, proteinexpression, or protein function of a Bcl-2 family protein. An inhibitormay selectively inhibit one, two, three, four, five, six or more membersof the Bcl-2 family proteins. In an embodiment, an inhibitor may affectnucleic acid or protein expression of a Bcl-2 family protein.Non-limiting examples of inhibitors that decrease nucleic acid andprotein expression may include histone deacetylase inhibitors such assodium butyrate and depsipeptide, synthetic cytotoxic retinoid such asfenretinide, and cyclin-dependent kinase inhibitors such asflavopiridol. Alternatively, an inhibitor may be an antisense molecule.For example, oblimersen sodium (G3139) is a Bcl-2 antisense that targetsBCL-2 mRNA. In another embodiment, an inhibitor may be a naturalinhibitor of Bcl-2 family interactions. For example, progidiosinmolecules (bypyrrole-containing natural products), such as GX15-070(obatoclax) may inhibit Bcl-2 family proteins such as Bcl-2, Bcl-xl,Bcl-w and Mcl-1. Additionally, the natural inhibitor gossypol (AT-101)and its derivatives, apogossypolone, TW37 and TM-1206, may inhibit Bcl-2family proteins such as Bcl-2, Bcl-xl, and Mcl-1. In still anotherembodiment, an inhibitor may be designed to bind the hydrophobic groveof anti-apoptotic Bcl-2 family proteins in place of BH3-only proteins(i.e., BH3-mimetics). As such, an inhibitor may be a small moleculeinhibitor of one or more anti-apoptotic proteins in the Bcl-2 family.For example, isoxazolidine-based small molecules that interact withBcl-2 and Bcl-xl, ABT-737 and ABT-263 (navitoclax) that bind Bcl-2,Bcl-xl, and Bcl-w. Non-limiting examples of other Bcl-2 familyinhibitors may include SAHB_(A), terphenyl, benzoylureas, A-385358,A-874009, A-1155463, A-1331852, apogossypolone, BM-1074, BM-1197,BXI-72, HA-14, antimycin A, ABT199, WEHI539, MIM-1, and BH₃Is. In aspecific embodiment, an inhibitor is a molecule similar to ABT-263. Inan exemplary embodiment, an inhibitor of one or more anti-apoptoticproteins in the Bcl-2 family is ABT-263 (navitoclax).

In an aspect, a composition of the invention further comprises ABT-263or an ABT-263 derivative. ABT-263 or ABT-263 derivatives may be modifiedto improve bioavailability, solubility, stability, handling properties,or a combination thereof, as compared to an unmodified version. Thus, inanother aspect, a composition of the invention may further comprisemodified ABT-263 or ABT-263 derivative. In still another aspect, acomposition of the invention further comprises a prodrug of ABT-263 oran ABT-263 derivative.

In an embodiment, the composition further comprises at least oneinhibitor of one or more anti-apoptotic proteins in the Bcl-2 family.For example, the composition may further comprise 1, 2, 3, 4 or 5 ormore inhibitors of one or more anti-apoptotic proteins in the Bcl-2family. Each Bcl-2 inhibitor of the composition may target the same ordifferent anti-apoptotic protein in the Bcl-2 family. In an embodiment,the composition may further comprise two inhibitors of one or moreanti-apoptotic proteins in the Bcl-2 family. In another embodiment, thecomposition may further comprise one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family.

Dosages of the Bcl-2 family inhibitor can vary between wide limits,depending upon the disease or disorder to be treated, the age andcondition of the subject to be treated. In an embodiment where thecomposition further comprising at least one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family is contacted with a sample,the concentration of the at least one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family may be from about 0.01 μM toabout 10 μM. Alternatively, the concentration of the at least oneinhibitor of one or more anti-apoptotic proteins in the Bcl-2 family maybe from about 0.01 μM to about 5 μM. For example, the concentration ofthe at least one inhibitor of one or more anti-apoptotic proteins in theBcl-2 family may be about 0.01, about 0.05, about 0.1, about 0.2, about0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9,about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8,about 9 or about 10 μM. Additionally, the concentration of the at leastone inhibitor of one or more anti-apoptotic proteins in the Bcl-2 familymay be greater than 10 μM. For example, the concentration of the atleast one inhibitor of one or more anti-apoptotic proteins in the Bcl-2family may be about 10, about 15, about 20, about 25, about 30, about35, about 40, about 45, about 50, about 55, about 60, about 65, about70, about 75, about 80, about 85, about 90, about 95 or about 100 μM.

In an embodiment where the composition further comprising at least oneinhibitor of one or more anti-apoptotic proteins in the Bcl-2 family isadministered to a subject, the dose of inhibitor may be from about 0.1mg/kg to about 500 mg/kg. For example, the dose of the least oneinhibitor of one or more anti-apoptotic proteins in the Bcl-2 family maybe about 0.1 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 5 mg/kg, about10 mg/kg, about 15 mg/kg, about 20 mg/kg, or about 25 mg/kg.Alternatively, the dose of the least one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family may be about 25 mg/kg, about50 mg/kg, about 75 mg/kg, about 100 mg/kg, about 125 mg/kg, about 150mg/kg, about 175 mg/kg, about 200 mg/kg, about 225 mg/kg, or about 250mg/kg. Additionally, the dose of the least one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family may be about 300 mg/kg,about 325 mg/kg, about 350 mg/kg, about 375 mg/kg, about 400 mg/kg,about 425 mg/kg, about 450 mg/kg, about 475 mg/kg or about 500 mg/kg.

II. Process of Making a Compound Comprising Formula (I) or Formula (II)

In an embodiment, a compound comprising Formula (I) may be synthesizedvia the reaction scheme depicted in Scheme 1.

By way of non-limiting example, Et₃N (about 1.15 eq) and pivaloylchloride (about 1.1 eq) may be added to a solution of acid (about 1 eq)in dry THF. The reaction mixture may be stirred for about 45-60 min forcompletion (monitored by TLC). The mixture may be filtered and thefiltrate may be used directly for the next step. About 0.88 eq of n-BuLimay be added to a stirred solution of 5,6-dihydropyridin-2(1H)-one (0.8eq) in dry THF under inert atmosphere at about −78° C. After stirringfor about 0.5 h, the mixed anhydride solution prepared from above may beadded and the resulting reaction mixture may be stirred for about 1 hbefore quenched with saturated NH₄Cl (about 0.5 mL). The mixture may beextracted with ethylacetate (10 mL×3). The organic phases may becombined, washed with sat. NaCl, dried over anhydrous Na₂SO₄, andevaporated to dryness under vacuum. The product may be purified purifiedby silica gel column chromatography. A skilled artisan would be able todetermine various changes to the reactions described above that wouldresult in various compounds comprising Formula (I).

In another embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 2.

By way of non-limiting example, a mixture of ethyl2-oxopiperidine-3-carboxylate and calcium chloride in ethanol may becooled to about 0° C. and treated with sodium borohydride. The resultingmixture may be allowed to warm to about room temperature and stirred forabout 6 hrs. The reaction may then quenched with acetone and the pH ofthe mixture may be adjusted to about pH to 3-4 with about 1.0 M HCl. Theresulting mixture may be extracted with methylene chloride, organicphases may be combined, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The product may be purified by silica gel columnchromatography. Then, cuprous iodine may be added to a stirred solutionof 3-(Hydroxymethyl)piperidin-2-one and dicyclohexylcarbodiimide (DCC)in toluene. The resulting mixture may be stirred under reflux for about1.5 hrs and cooled to room temperature. Water may be added and themixture stirred overnight. Ethylacetate may be added and the mixture maybe filtered. The aqueous phase may be extracted with ethyl acetate andthe combined organic phases may be washed with brine, dried over Na₂SO₄,filtered, and evaporated to dryness under vacuum. The product may bepurified by silica gel column chromatography. Next, a solution of3-Methylene-2-piperidinone in THF may be cooled to about −78° C., n-BuLimay be added slowly. The resulting mixture may be allowed to stir atabout −78° C. for about 1 hr. A solution of(E)-3-(3,4,5-trimethoxyphenyl)acryloyl chloride in THF may then be addeddropwise. After addition, the mixture may be stirred under about −78° C.for about 1 hr. The reaction may then quenched with aqueous ammoniumchloride solution and may be extracted with ethylacetate. The combinedorganic phases may be washed with brine, dried over Na₂SO₄, filtered,and evaporated to dryness under vacuum. The product may be purified bysilica gel column chromatography. A skilled artisan would be able todetermine various changes to the reactions described above that wouldresult in various compounds comprising Formula (II).

In still another embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 3.

By way of non-limiting example, ethyl 2-oxopyrrolidine-3-carboxylate maybe added to a suspension of NaBH₄ and CaCl₂ in ethanol under about 0° C.and the resulting mixture may stirred overnight at about roomtemperature. The reaction may be quenched with about 1M HCl solution,then extracted with CH₂Cl₂/MeOH(10/1) about 10 to 20 times, the combinedorganic phases may be dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The product may be purified by silica gel columnchromatography. Next, 3-(Hydroxymethyl)pyrrolidin-2-one may be treatedwith CuI and DCC in toluene under reflux for about 3 h. The reactionmixture may be cooled to about room temperature and quenched withseveral drops of water, followed by filtration, the filtrate may beconcentrated and purified through column. Then,3-Methylenepyrrolidin-2-one may be dissolved in THF and cooled to about−78° C., and n-BuLi may be added dropwise under N₂ protection, theresulting mixture may be stirred under about −78° C. for about 1 h.(E)-t-butyl 3-(3,4,5-trimethoxyphenyl)acrylate (about 1.5 eq) may beadded. After stirring at about −78° C. for about 1 h, the reaction maybe quenched with NH₄Cl solution, extracted with EtOAc about 3 times, thecombined organic phases may be washed with brine and dried over Na₂SO₄,filtered, and evaporated to dryness under vacuum. The residue may bepurified by flash column chromatography. A skilled artisan would be ableto determine various changes to the reactions described above that wouldresult in various compounds comprising Formula (II).

In still yet another embodiment, a compound comprising Formula (II) maybe synthesized via the reaction scheme depicted in Scheme 4.

By way of non-limiting example, LHMDS may be added to a solution of(E)-1-(3-(3,4,5-trimethoxyphenyl)acryloyl)piperidin-2-one in THF atabout −78° C. The resulting solution may be stirred at this temperaturefor about 1 h. Aromatic aldehyde (about 1 eq; dissolved in small amountof THF) may be added dropwise. After stirring at about −78° C. for aboutan additional hour, ammonium chloride solution may be added to quenchthe reaction, the mixture may be extracted with EtOAc about 3 times. Thecombined organic phases may be washed with water and brine, dried overNa₂SO₄, filtered, and evaporated to dryness under vacuum. The residuemay be purified by flash column chromatography to afford thecorresponding hydroxyl intermediate. Next, methanesulfonyl chloride(about 1.2 eq) may be added to a solution of the above intermediate andTEA (about 1.5 eq) in toluene under ice bath. After stirring at aboutroom temperature for about 1 h, additional about 2.5 eq TEA may beadded. The resulting mixture may be heated to reflux for about 5.5 h.Upon cooled to about room temperature, the reaction may be quenched withwater and extracted with ethylacetate. The combined organic fractionsmay be washed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The residue may be purified by flash columnchromatography to afford the desired product. A skilled artisan would beable to determine various changes to the reactions described above thatwould result in various compounds comprising Formula (II).

In a different embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 5.

By way of non-limiting example, LHDMS may be added to a solution oftert-butyl 2-oxopiperidine-1-carboxylate in THF at about −78° C. underN₂ protection. After stirring at this temperature for about 1 h,isobutyraldehyde may be added dropwise. The resulting mixture may beallowed to stir for about an additional hour before quenched by slowlyadding saturated NH₄Cl solution, then diluted with EtOAc and the organicfraction may be washed with brine, dried over Na₂SO₄, filtered, andevaporated to dryness under vacuum. The residue may be purified by flashcolumn chromatography to afford the desired product. Next,methanesulfonyl chloride may be added to a solution of t-butyl3-(1-hydroxy-2-methylpropyl)-2-oxopiperidine-1-carboxylate and TEA intoluene at about 0° C. After stirring at about room temperature forabout 1 h, additional TEA may then be added, the resulting mixture maybe heated to reflux and stirred under reflux for about 6 h. Aftercooling down to about room temperature, the reaction may be quenchedwith water and extracted with EtOAc. The combined organic fractions maybe washed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The residue may be purified by flash columnchromatography to afford both a E isomer and Z isomer product. Next, TFAmay be added to a solution of t-butyl3-(2-methylpropylidene)-2-oxopiperidine-1-carboxylate (Z or E isomer) inCH₂Cl₂ at about 0° C. The resulting mixture may be allowed to stir atabout 0° C. for about 2 h. The reaction may then be quenched withsaturated sodium bicarbonate under ice bath and extracted with CH₂Cl₂.The combined organic fractions may be washed with brine, dried overNa₂SO₄, filtered, and evaporated to dryness under vacuum. The residuemay be purified by flash column chromatography to afford thecorresponding Z or E isomer. Next, n-BuLi may be added to a solution of3-(2-methylpropylidene) piperidin-2-one (Z or E isomer) in THF at about−78° C. under N₂ protection. After stirring at about −78° C. for about 1h, (E)-tert-butyl 3-(3,4,5-trimethoxyphenyl)acrylate may be added. Theresulting mixture may be stirred at about −78° C. for about 45 min andthen quenched with saturated NH₄Cl water solution. The whole mixture maybe extracted with EA several times. The combined organic phase may bewashed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The residue may be purified by flash columnchromatography to afford the corresponding Z or E isomer. A skilledartisan would be able to determine various changes to the reactionsdescribed above that would result in various compounds comprisingFormula (II).

In another embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 6.

By way of non-limiting example, n-BuLi may be added to a solution of3-methylenepiperidin-2-one in THF at about −78° C. After stirring atthis temperature for about 1 h, (E)-2-phenylethenesulfonyl chloride(dissolved in small amount of THF) may be added. The resulting mixturemay be stirred for about another 1 h and then quenched with saturatedNH₄Cl water solution. The whole mixture may be extracted with EA severaltimes. The combined organic phase may be washed with brine, dried overNa₂SO₄, filtered, and evaporated to dryness under vacuum. The residuemay purified by flash column chromatography to afford the desiredproduct. A skilled artisan would be able to determine various changes tothe reactions described above that would result in various compoundscomprising Formula (II).

In another embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 7.

By way of non-limiting example, Boc₂O may be added to a solution ofε-Caprolactam, DMAP and Et₃N in CH₂Cl₂ at about 0° C. The resultingmixture may be stirred at about room temperature overnight. Solvent maybe evaporated giving a residue which may be purified. Next, LHMDS may beadded to a solution of tert-butyl 2-oxoazepane-1-carboxylate in THF atabout −78° C. The resulting mixture may be stirred at this temperaturefor about 45 min and methyl carbonochloridate may then be added. Thewhole mixture may be stirred for about an additional 1 h and quenchedwith saturated NH₄Cl water solution, followed by extraction with EAseveral times. The combined organic phase may be washed with brine anddried over anhydrous sodium sulfate. Solvent may be evaporated giving aresidue which may be purified. Next, paraformaldehyde may be added to asuspension of 1-tert-butyl 3-methyl 2-oxoazepane-1,3-dicarboxylate,18-crown-6, K₂CO₃ in toluene. The resulting mixture may be stirred underreflux for about 4-5 hours and then cooled to about room temperature andwater may be added. The whole mixture may be extracted with EA about 3times. The combined organic phase may be washed with brine and driedover anhydrous sodium sulfate. Solvent may be evaporated giving aresidue which may be purified. Next, TFA may be added to a solution oftert-butyl 3-methylene-2-oxoazepane-1-carboxylate in CH₂Cl₂ at 0° C. Theresulting solution may be stirred at this temperature for about 2 h.Saturated sodium bicarbonate solution may be added until pH larger thanabout 7. The mixture may then be extracted with CH₂Cl₂ about 5-10 timesand the combined organic phase may be washed with brine and dried overanhydrous sodium sulfate. Solvent may be evaporated giving a residuewhich may be purified. Next, n-BuLi may be added to a solution of3-methyleneazepan-2-one in THF at about −78° C. After stirring at thistemperature for about 1 h, (E)-tert-butyl3-(3,4,5-trimethoxyphenyl)acrylate may then be added. The resultingmixture may be stirred at about −78° C. for about an additional 1 h andthen quenched with saturated NH₄Cl solution. The whole mixture may thenbe extracted with EA several times and the combined organic phase may bewashed with brine and dried over anhydrous sodium sulfate. Solvent maybe evaporated giving a residue which may be purified chromatographicallyto get pure product. A skilled artisan would be able to determinevarious changes to the reactions described above that would result invarious compounds comprising Formula (II).

In still another embodiment, a compound comprising Formula (II) may besynthesized via the reaction scheme depicted in Scheme 8.

By way of non-limiting example, TFAA may be added to a solution ofazepan-2-onein toluene at about 0° C. After stirring under ice bath forabout 1 h, the solvent may be removed to give a residue. The residue maybe mixed with 2-chlorobenzaldehyde and charged to a suspension of t-BuOKin THF at about 0° C. The resulting mixture may then be heated to about50-60° C. and stirred at this temperature for about 1.5 h. The reactionmixture may be concentrated under vacuum and water may then be added.The resulting mixture may be extracted with EA about 3 times. Thecombined organic phase may be washed with brine and dried over anhydroussodium sulfate. Solvent may be evaporated giving a residue which may bepurified. Next, n-BuLi may be added to a solution of(E)-3-(2-chlorobenzylidene)azepan-2-one in THF at about −78° C. Afterstirring at about −78° C. for about 1 h, (E)-tert-butyl3-(3,4,5-trimethoxyphenyl)acrylate may be added and the resultingmixture may be stirred at about −78° C. for about an additional 1 h. Thereaction may be quenched with saturated NH₄Cl solution and thenextracted with EA about 3 times. The combined organic phase may bewashed with brine and dried over anhydrous sodium sulfate. Solvent maybe evaporated giving a residue which may be purified to get pureproduct. A skilled artisan would be able to determine various changes tothe reactions described above that would result in various compoundscomprising Formula (II).III. Methods

The present disclosure encompasses a method of selectively killing oneor more senescent cells in a sample, the method comprising contacting acomposition comprising an effective amount of PL, a PL derivative, acompound of Formula (I) or a compound of Formula (II) with the sample.In another aspect, the present disclosure encompasses a method ofselectively killing one or more senescent cells in a subject in needthereof, the method comprising administering to the subject acomposition comprising a therapeutically effective amount PL, a PLderivative, a compound of Formula (I) or a compound of Formula (II).

By selectively killing one or more senescent cells is meant acomposition of the invention does not appreciably kill non-senescentcells at the same concentration. Accordingly, the median lethal dose orLD50 of the inhibitor in non-senescent cells may be about 5 to about 50times higher than the LD50 of the inhibitor in senescent cells. As usedherein, the LD50 is the concentration of inhibitor required to kill halfthe cells in the cell sample. For example, the LD50 of the inhibitor innon-senescent cells may be greater than about 5, about 6, about 7, about8, about 9 or about 10 times higher than the LD50 of the inhibitor insenescent cells. Alternatively, the LD50 of the inhibitor innon-senescent cells may be greater than about 10, about 15, about 20,about 25, about 30, about 35, about 40, about 45, or about 50 timeshigher than the LD50 of the inhibitor in senescent cells. Additionally,the LD50 of the inhibitor in non-senescent cells may be greater than 50times higher than the LD50 of the inhibitor in senescent cells. In aspecific embodiment, the LD50 of the inhibitor in non-senescent cells isgreater than 10 times higher than the LD50 of the inhibitor in senescentcells. In another specific embodiment, the LD50 of the inhibitor innon-senescent cells is greater than 20 times higher than the LD50 of theinhibitor in senescent cells.

The progression from an actively dividing cell to a metabolicallyactive, non-dividing cell is termed “senescence” or “cellularsenescence.” As used herein, the terms “senescence” and “cellularsenescence” may be used interchangeably. The term “senescence” alsorefers to the state into which cells enter after multiple rounds ofdivision and, as a result of cellular pathways, future cell division isprevented from occurring even though the cell remains metabolicallyactive. Senescent cells may differ from their pre-senescent counterpartsin one or more of the following ways: 1) they arrest growth and cannotbe stimulated to reenter the cell cycle by physiological mitogens; 2)they become resistant to apoptotic cell death; and/or 3) they acquirealtered differentiated functions.

In contrast to cancer cells which grow and divide uncontrollably, theability of most differentiated eukaryotic cells to proliferate isfinite. Stated another way, normal cells have an intrinsicallydetermined limit to the number of cell divisions through which they canproceed. This phenomenon has been termed “replicative cellularsenescence” and is an intrinsic anticancer mechanism that limits acell's proliferative ability, thereby preventing neoplastictransformation. Another form of senescence is “premature cellularsenescence.” Premature cellular senescence, like replicative cellularsenescence, is a terminal fate of mitotic cells, characterized bypermanent cell cycle arrest. Unlike replicative cellular senescence,however, premature cellular senescence does not require telomeredeterioration and can be induced by a variety of stressors including,but not limited to, ultraviolet light, reactive oxygen species,chemotherapeutics, environmental toxin, cigarette smoking, ionizingradiation, distortion of chromatin structure, excessive mitogenicsignaling, and oncogenic mutations. Still another form of senescence istherapy-induced senescence (TIS) which refers to the phenomenon of asubset of tumor cells being forced into a senescent state by therapeuticagents. TIS is known to develop because of certain treatments, includingradiotherapy and chemotherapy.

The number of senescent cells in various organs and tissues of a subjectincreases with age. The accumulation of senescent cells may drive thedeterioration that underlies aging and age-related diseases. Forexample, the accumulation of senescent cells in aged tissue maycontribute to age-associated tissue dysfunction, reduced regenerativecapacity, and disease. In this context, senescence is considereddeleterious because it contributes to decrements in tissue renewal andfunction. As a non-limiting example, an aged tissue may lack the abilityto respond to stress when proliferation is required thereby resulting inthe reduced fitness seen with aging. A key component of this model isthat substantial numbers of senescent cells should be present in tissueswith aging, without, or prior to, pathology.

(a) Senescent Cells

A senescent cell may be a cell that ceases to divide but remainsmetabolically active. The non-dividing cells may remain viable for manyweeks, but fail to grow/replicate DNA despite the presence of amplespace, nutrients and growth factors in the medium. Thus, the senescencegrowth arrest is essentially permanent because senescent cells cannot bestimulated to proliferate by known physiological stimuli. Further, asenescent cell of the invention may be resistant to certain apoptoticsignals and may acquire widespread changes in gene expression. Theresistance to apoptosis may explain the increase in senescent cells withage. Manipulation of pro- and anti-apoptotic proteins may cause cellsthat are destined to die by apoptosis to senesce and, conversely, causecells that are destined to senesce to undergo apoptosis.

A senescent cell of the invention may be senescent due to replicativecellular senescence, premature cellular senescence or therapy-inducedsenescence. Senescent cells that are senescent due to replication mayhave undergone greater than 60 population doublings. Alternatively,senescent cells that are senescent due to replication may have undergonegreater than 40, greater than 50, greater than 60, greater than 70 orgreater than 80 population doublings. A senescent cell that isprematurely cellular senescent may be induced by, but not limited to,ultraviolet light, reactive oxygen species, chemotherapeutics,environmental toxin, cigarette smoking, ionizing radiation, distortionof chromatin structure, excessive mitogenic signaling, and oncogenicmutations. In a specific embodiment, premature cellular senescence maybe induced by ionizing radiation (IR). In another specific embodiment,premature cellular senescence may also be induced by ectopictransfection with Ras oncogene. A senescent cell that is therapy-inducedsenescent may have been exposed to DNA-damaging therapy.

A senescent cell of the invention may generally be a eurkaryotic cell.Non-limiting examples of senescent cells may include, but are notlimited to, mammary epithelial cells, keratinocytes, cardiac myocytes,chondrocytes, endothelial cells (large vessels), endothelial cells(microvascular), epithelial cells, fibroblasts, follicle dermal papillacells, hepatocytes, melanocytes, osteoblasts, preadipocytes, primarycells of the immune system, skeletal muscle cells, smooth muscle cells,adipocytes, neurons, glial cells, contractile cells, exocrine secretoryepithelial cells, extracellular matrix cells, hormone secreting cells,keratinizing epithelial cells, islet cells, lens cells, mesenchymal stemcells, pancreatic acinar cells, paneth cells of the small intestine,primary cells of hemopoietic linage, primary cells of the nervoussystem, sense organ and peripheral neuron supporting cells, wetstratified barrier epithelial cells and stem cells. In a specificembodiment, the stem cells are adult stem cells. Adult stem cells arestem cells which maintain and repair the tissue in which they are foundand are generally referred to by their tissue of origin. Non-limitingexamples of adult stem cells include muscle stem cells, hematopoieticstem cells, heart stem cells, neural stem cells, mesenchymal stem cells,intestinal stem cells, skin stem cells, adipose-derived stem cells,endothelial stem cells, and dental pulp stem cells. In a specificembodiment, a senescent cell of the invention is a fibroblast. Inanother specific embodiment, a senescent cell may be a hematopoieticstem cell.

Further, a senescent cell of the invention may be found in renewabletissues, including the vasculature, hematopoietic system, epithelialorgans and the stroma. A senescent cell of the invention may also befound at sites of aging or chronic age-related pathology, such asosteoarthritis and atherosclerosis. Further, a senescent cell of theinvention may be associated with benign dysplastic or preneoplasticlesions and benign prostatic hyperplasia. In an embodiment, a senescentcell of the invention may be found in normal and tumor tissues followingDNA-damaging therapy. In a specific embodiment, a senescent cell may befound at a site of aging or age-related pathology.

An age-related pathology may include any disease or condition which isfully or partially mediated by the induction or maintenance of anon-proliferating or senescent state in a cell or a population of cellsin a subject. Non-limiting examples include age-related tissue or organdecline which may lack visible indication of pathology, or overtpathology such as a degenerative disease or a function-decreasingdisorder. For example, Alzheimer's disease, Parkinson's disease,cataracts, macular degeneration, glaucoma, atherosclerosis, acutecoronary syndrome, myocardial infarction, stroke, hypertension,idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonarydisease (COPD), osteoarthritis, type 2 diabetes, obesity, fatdysfunction, coronary artery disease, cerebrovascular disease,periodontal disease, and cancer treatment-related disability such asatrophy and fibrosis in various tissues, brain and heart injury, andtherapy-related myelodysplastic syndromes. Additionally, an age-relatedpathology may include an accelerated aging disease such as progeroidsyndromes (i.e. Hutchinson-Gilford progeria syndrome, Werner syndrome,Bloom syndrome, Rothmund-Thomson Syndrome, Cockayne syndrome, xerodermapigmentosum, trichothiodystrophy, combined xerodermapigmentosum-Cockayne syndrome, restrictive dermopathy), ataxiatelangiectasia, Fanconi anemia, Friedreich's ataxia, dyskeratosiscongenital, aplastic anemia, IPF, and others. A method of identifying anage-related disease or condition as described herein may includedetecting the presence of senescent cells.

(b) Detecting Senescent Cells

In an aspect, a method of the invention may comprise detecting senescentcells. Senescent cells may be detected in vivo or in vitro. Suitablemarkers for detecting senescent cells in vitro and in vivo are known inthe art. For example, methods to detect senescent cells may include, butare not limited to, detecting lack of DNA replication by incorporationof a DNA-staining reagent (e.g. 5-bromodeoxyuridine (BrdU),³H-thymidine), immunostaining for proteins such as proliferating cellnuclear antigen (PCNA) and Ki-67, histochemical staining forsenescence-associated β-galactosidase (SA-β-gal), detecting expressionof p16, p19, Pai1, Igfbp2, IL-6, Mmp13, Nrg1, differentiatedembryo-chondrocyte expressed-1 (DEC1), p15 (a CDK1) and decoy deathreceptor-2 (DCR2), detecting cytological markers such assenescence-associated heterochromatin foci (SAHFs) andsenescence-associated DNA-damage foci (SDFs). SAHFs may be detected bythe preferential binding of DNA dyes, such as4′,6-diamidino-2-phenylindole (DAPI), and the presence of certainheterochromatin-associated histone modifications (for example, H3 Lys9methylation) and proteins (for example, heterochromatin protein-1(HP1)). Additionally, senescent cells may be detected as described inU.S. Pat. No. 5,491,069 and US Patent Application No. 2010/0086941. Incertain embodiments, senescent cells are detected by histochemicalstaining for SA-β-gal.

In certain embodiments, one or more senescent cells are detected in asample. A sample may be a cell sample, a tissue sample, or a biopsy froma subject. Generally speaking, a sample may be dependent on theage-related pathology. For instance, a sample may be tissue biopsymaterial. As such, a tissue sample may be from esophagus, stomach,liver, gallbladder, pancreas, adrenal glands, bladder, gallbladder,large intestine, small intestine, kidneys, liver, pancreas, colon,stomach, thymus, spleen, brain, spinal cord, nerves, adipose tissue,heart, lungs, eyes, corneal, skin or islet tissue or organs. In aspecific embodiment, a tissue sample may be from lung, skeletal muscle,and brain. In another specific embodiment, a tissue sample may be fromliver and heart. Alternatively, a sample may be a cell sample. As such,a cell sample may be oocytes and/or spermatozoa, mesenchymal stem cells,adipocytes, central nervous system neurons and glial cells, contractilecells, exocrine secretory epithelial cells, extracellular matrix cells,hormone secreting cells, keratinizing epithelial cells, islet cells,kidney cells, lens cells, pancreatic acinar cells, paneth cells of smallintestine, primary cells of hemopoietic lineage, primary cells of thenervous system, sense organ and peripheral neuron supporting cells orwet stratified barrier epithelial cells. Detection of senescent cellsmay be used to assess the replicative history of tissues, therebyproviding a method for evaluation of the physiological, in contrast tothe chronological age of the tissue.

The number of senescent cells may increase with age. The number ofsenescent cells in a tissue or sample may be from less than 1% togreater than 15%. In an embodiment, the number of senescent cells in atissue or sample may be less than 1%, less than 2%, less than 3%, lessthan 4%, or less than 5%. In another embodiment, the number of senescentcells in a tissue or sample may be about 5%, about 6%, about 7%, about8%, about 9%, or about 10%. In still another embodiment, the number ofsenescent cells in a tissue or sample may be greater than 10%, greaterthan 11%, greater than 12%, greater than 13%, greater than 14%, orgreater than 15%.

(c) Measuring Cell Death

In an aspect, a method of the invention may comprise measuring celldeath of senescent cells. Methods of measuring cell death are known inthe art. For example, cell death may be measured by Giemsa staining,trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) doublestaining for fluorescence microscopy and flow cytometry, propidiumiodide (PI) staining, annexin V assay, TUNEL assay, DNA ladder, LDHactivity, and MTT assay. In a preferred embodiment, cell death is due toinduction of apoptosis. Cell death due to induction of apoptosis may bemeasured by observation of morphological characteristics including cellshrinkage, cytoplasmic condensation, chromatin segregation andcondensation, membrane blebbing, and the formation of membrane-boundapoptotic bodies. Cell death due to induction of apoptosis may bemeasured by observation of biochemical hallmarks includinginternucleosomal DNA cleavage into oligonucleosome-length fragments.Traditional cell-based methods of measuring cell death due to inductionof apoptosis include light and electron microscopy, vital dyes, andnuclear stains. Biochemical methods include DNA laddering, lactatedehydrogenase enzyme release, and MTT/XTT enzyme activity. Additionally,terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick endlabeling of DNA fragments (TUNEL) and in situ end labeling (ISEL)techniques are used, which when used in conjunction with standard flowcytometric staining methods yield informative data relating cell deathto various cellular parameters, including cell cycle and cell phenotype.See Loo and Rillema, Methods Cell Biol. 1998; 57:251-64, which isincorporated herein by reference, for a review of these methods. In anexemplary embodiment, cell death due to apoptosis may be measured by thereduction of procaspase-3. Caspase-3 has been implicated as an“effector” caspase associated with the initiation of the “death cascade”and is therefore an important marker of the cell's entry point into theapoptotic signaling pathway. Caspase-3 is activated by the upstreamcaspase-8 and caspase-9, and since it serves as a convergence point fordifferent signaling pathways, it is well suited as a read-out in anapoptosis assay.

The results of these methods may be used to determine the percentage ofviable cells. In a preferred embodiment, cell death may be measured as areduction in viable cells. Since a composition of the inventionselectively kills senescent cells, a reduction in viable cells isindicative of a reduction in senescent cells. As described in SectionIII(b), the number of senescent cells in a sample may be from less than1% to greater than 15%. As such, a reduction in viable cells followingadministration of an inhibitor of the invention may be greater than 15%to less than 1%. For example, the reduction in viable cells may be lessthan 1%, less than 2%, less than 3%, less than 4%, or less than 5%.Alternatively, the reduction in viable cells may be about 5%, about 6%,about 7%, about 8%, about 9%, or about 10%. Additionally, the reductionin viable cells may be greater than 10%, greater than 11%, greater than12%, greater than 13%, greater than 14%, or greater than 15%.

(d) Administration

In certain aspects, a therapeutically effective amount of a compositionof the invention may be administered to a subject. Administration isperformed using standard effective techniques, including peripherally(i.e. not by administration into the central nervous system) or locallyto the central nervous system. Peripheral administration includes but isnot limited to oral, inhalation, intravenous, intraperitoneal,intra-articular, subcutaneous, pulmonary, transdermal, intramuscular,intranasal, buccal, sublingual, or suppository administration. Localadministration, including directly into the central nervous system (CNS)includes but is not limited to via a lumbar, intraventricular orintraparenchymal catheter or using a surgically implanted controlledrelease formulation. The route of administration may be dictated by thedisease or condition to be treated. For example, if the disease orcondition is COPD or IPF, the composition may be administered viainhalation. Alternatively, is the disease or condition isosteoarthritis, the composition may be administered via intra-articularinvention. It is within the skill of one in the art, to determine theroute of administration based on the disease or condition to be treated.In a specific embodiment, a composition of the invention is administeredorally.

Pharmaceutical compositions for effective administration aredeliberately designed to be appropriate for the selected mode ofadministration, and pharmaceutically acceptable excipients such ascompatible dispersing agents, buffers, surfactants, preservatives,solubilizing agents, isotonicity agents, stabilizing agents and the likeare used as appropriate. Remington's Pharmaceutical Sciences, MackPublishing Co., Easton Pa., 16 Ed ISBN: 0-912734-04-3, latest edition,incorporated herein by reference in its entirety, provides a compendiumof formulation techniques as are generally known to practitioners.

For therapeutic applications, a therapeutically effective amount of acomposition of the invention is administered to a subject. A“therapeutically effective amount” is an amount of the therapeuticcomposition sufficient to produce a measurable response (e.g., celldeath of senescent cells, an anti-aging response, an improvement insymptoms associated with a degenerative disease, or an improvement insymptoms associated with a function-decreasing disorder). Actual dosagelevels of active ingredients in a therapeutic composition of theinvention can be varied so as to administer an amount of the activecompound(s) that is effective to achieve the desired therapeuticresponse for a particular subject. The selected dosage level will dependupon a variety of factors including the activity of the therapeuticcomposition, formulation, the route of administration, combination withother drugs or treatments, age, the age-related disease or condition,the degenerative disease, the function-decreasing disorder, thesymptoms, and the physical condition and prior medical history of thesubject being treated. In some embodiments, a minimal dose isadministered, and dose is escalated in the absence of dose-limitingtoxicity. Determination and adjustment of a therapeutically effectivedose, as well as evaluation of when and how to make such adjustments,are known to those of ordinary skill in the art of medicine.

The frequency of dosing may be daily or once, twice, three times or moreper week or per month, as needed as to effectively treat the symptoms.The timing of administration of the treatment relative to the diseaseitself and duration of treatment will be determined by the circumstancessurrounding the case. Treatment could begin immediately, such as at thesite of the injury as administered by emergency medical personnel.Treatment could begin in a hospital or clinic itself, or at a later timeafter discharge from the hospital or after being seen in an outpatientclinic. Duration of treatment could range from a single doseadministered on a one-time basis to a life-long course of therapeutictreatments.

Typical dosage levels can be determined and optimized using standardclinical techniques and will be dependent on the mode of administration.

(e) Subject

A subject may be a rodent, a human, a livestock animal, a companionanimal, or a zoological animal. In one embodiment, the subject may be arodent, e.g. a mouse, a rat, a guinea pig, etc. In another embodiment,the subject may be a livestock animal. Non-limiting examples of suitablelivestock animals may include pigs, cows, horses, goats, sheep, llamasand alpacas. In still another embodiment, the subject may be a companionanimal. Non-limiting examples of companion animals may include pets suchas dogs, cats, rabbits, and birds. In yet another embodiment, thesubject may be a zoological animal. As used herein, a “zoologicalanimal” refers to an animal that may be found in a zoo. Such animals mayinclude non-human primates, large cats, wolves, and bears. In apreferred embodiment, the subject is a human.

The human subject may be of any age. However, since senescent cells arenormally associated with aging, a human subject may be an older humansubject. In some embodiments, the human subject may be about 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 years of age or older. Insome preferred embodiments, the human subject is 30 years of age orolder. In other preferred embodiments, the human subject is 40 years ofage or older. In other preferred embodiments, the human subject is 45years of age or older. In yet other preferred embodiments, the humansubject is 50 years of age or older. In still other preferredembodiments, the human subject is 55 years of age or older. In otherpreferred embodiments, the human subject is 60 years of age or older. Inyet other preferred embodiments, the human subject is 65 years of age orolder. In still other preferred embodiments, the human subject is 70years of age or older. In other preferred embodiments, the human subjectis 75 years of age or older. In still other preferred embodiments, thehuman subject is 80 years of age or older. In yet other preferredembodiments, the human subject is 85 years of age or older. In stillother preferred embodiments, the human subject is 90 years of age orolder.

Additionally, a subject in need thereof may be a subject suffering froman age-related disease or condition as described below.

(f) Aging and Age-Related Diseases

It has been demonstrated that senescent cells drive age-relatedpathologies and that selective elimination of these cells can prevent ordelay age-related deterioration. Thus, senescent cells may betherapeutic targets in the treatment of aging and age-related disease.As such, removal of senescent cells may delay tissue dysfunction andextend health span. Clearance of senescent cells is expected to improvetissue milieu, thereby improving the function of the remainingnon-senescent cells.

The present disclosure provides a method for delaying at least onefeature of aging in a subject, the method comprising administering acomposition comprising a therapeutically effective amount of PL, a PLderivative, a compound of Formula (I) or a compound of Formula (II) to asubject. As used herein, “a feature of aging” may include, but is notlimited to, systemic decline of the immune system, muscle atrophy anddecreased muscle strength, decreased skin elasticity, delayed woundhealing, retinal atrophy, reduced lens transparency, reduced hearing,osteoporosis, sarcopenia, hair graying, skin wrinkling, poor vision,frailty, and cognitive impairment.

In an aspect, a composition of in the invention selectively killssenescent cells. In this way, targeting senescent cells during thecourse of aging may be a preventative strategy. Accordingly,administration of a composition comprising a therapeutically effectiveamount of PL, a PL derivative, a compound of Formula (I) or a compoundof Formula (II) to a subject may prevent comorbidity and delay mortalityin an older subject. Further, selective killing of senescent cells mayboost the immune system, extend the health span, and improve the qualityof life in a subject. Additionally, selective killing of senescent cellsmay delay sarcopenia. Sarcopenia is the degenerative loss of skeletalmuscle mass, quality, and strength associated with aging. As such, adelay in sarcopenia may reduce frailty, reduce risk of falling, reducefractures, and reduce functional disability in a subject. Furthermore,selective killing of senescent cells may delay aging of the skin. Agedskin has increased wrinkles, decreased immune barrier function andincreased susceptibility to skin cancer and trauma. As such, selectivekilling of senescent cells may delay skin wrinkling, delay the onset ofdecreased immune barrier function and decrease susceptibility to skincancer and trauma in a subject. Selective killing of senescent cells mayalso delay the onset of retinal atrophy and reduced lens transparency asmeasured by vision tests.

Methods of measuring aging are known in the art. For example, aging maybe measured in the bone by incident non-vertebral fractures, incidenthip fractures, incident total fractures, incident vertebral fractures,incident repeat fractures, functional recovery after fracture, bonemineral density decrease at the lumbar spine and hip, rate of kneebuckling, NSAID use, number of joints with pain, and osteoarthritis.Aging may also be measured in the muscle by functional decline, rate offalls, reaction time and grip strength, muscle mass decrease at upperand lower extremities, and dual tasking 10-meter gait speed. Further,aging may be measured in the cardiovascular system by systolic anddiastolic blood pressure change, incident hypertension, majorcardiovascular events such as myocardial infarction, stroke, congestiveheart disease, and cardiovascular mortality. Additionally, aging may bemeasured in the brain by cognitive decline, incident depression, andincident dementia. Also, aging may be measured in the immune system byrate of infection, rate of upper respiratory infections, rate offlu-like illness, incident severe infections that lead to hospitaladmission, incident cancer, rate of implant infections, and rate ofgastrointestinal infections. Other indications of aging may include, butnot limited to, decline in oral health, tooth loss, rate of GI symptoms,change in fasting glucose and/or insulin levels, body composition,decline in kidney function, quality of life, incident disabilityregarding activities of daily living, and incident nursing homeadmission. Methods of measuring skin aging are known in the art and mayinclude trans-epidermal water loss (TEWL), skin hydration, skinelasticity, area ratio analysis of crow's feet, sensitivity, radiance,roughness, spots, laxity, skin tone homogeneity, softness, and relief(variations in depth).

The present disclosure also provides a method of treating an age-relateddisease or condition, the method comprising administering a compositioncomprising a therapeutically effective amount of PL, a PL derivative, acompound of Formula (I) or a compound of Formula (II) to a subject inneed thereof, provided the age-related disease or condition is notcancer. As used herein, “age-related disease or condition” may include,but is not limited to, a degenerative disease or a function-decreasingdisorder such as Alzheimer's disease, Parkinson's disease, cataracts,macular degeneration, glaucoma, atherosclerosis, acute coronarysyndrome, myocardial infarction, stroke, hypertension, idiopathicpulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD),osteoarthritis, type 2 diabetes, obesity, fat dysfunction, coronaryartery disease, cerebrovascular disease, periodontal disease, cancertreatment-related disability such as atrophy and fibrosis in varioustissues, brain and heart injury, and therapy-related myelodysplasticsyndromes, and diseases associated with accelerated aging and/or defectsin DNA damage repair and telomere maintenance such as progeroidsyndromes (i.e. Hutchinson-Gilford progeria syndrome, Werner syndrome,Bloom syndrome, Rothmund-Thomson Syndrome, Cockayne syndrome, xerodermapigmentosum, trichothiodystrophy, combined xerodermapigmentosum-Cockayne syndrome, restrictive dermopathy), ataxiatelangiectasia, Fanconi anemia, Friedreich's ataxia, dyskeratosiscongenital, aplastic anemia, IPF, and others. Methods of diagnosing andidentifying an age-related disease or condition are known in the art.

The present disclosure also provides a method of killing therapy-inducedsenescent cells. The method comprises administering a compositioncomprising a therapeutically effective amount of PL or a compoundcomprising Formula (I) to a subject that has received DNA-damagingtherapy and killing therapy induced-senescent cells in normal and tumortissues following DNA-damaging therapy.

Non-limiting examples of DNA-damaging therapy may include γ-irradiation,alkylating agents such as nitrogen mustards (chlorambucil,cyclophosphamide, ifosfamide, melphalan), nitrosoureas (streptozocin,carmustine, lomustine), alkyl sulfonates (busulfan), triazines(dacarbazine, temozolomide) and ethylenimines (thiotepa, altretamine),platinum drugs such as cisplatin, carboplatin, oxalaplatin,antimetabolites such as 5-fluorouracil, 6-mercaptopurine, capecitabine,cladribine. clofarabine, cytarabine, floxuridine, fludarabine,gemcitabine, hydroxyurea, methotrexate, pemetrexed, pentostatin,thioguanine, anthracyclines such as daunorubicin, doxorubicin,epirubicin, idarubicin, anti-tumor antibiotics such as actinomycin-D,bleomycin, mitomycin-C, mitoxantrone, topoisomerase inhibitors such astopoisomerase I inhibitors (topotecan, irinotecan) and topoisomerase IIinhibitors (etoposide, teniposide, mitoxantrone), mitotic inhibitorssuch as taxanes (paclitaxel, docetaxel), epothilones (ixabepilone),vinca alkaloids (vinblastine, vincristine, vinorelbine) andestramustine.

Based on the observation that ionizing radiation and variouschemotherapeutic agents elicit a marked senescence response in vivo,therapy-induced senescent cells may be a cause of long-termramifications after DNA-damaging therapy, such as cancer therapy. Assuch, the systemic accumulation of therapy-induced senescent cells maydrive accelerated physical decline in cancer survivors. Acceleratedphysical decline may also be referred to as accelerated aging.Accordingly, once a tumor is removed by systemic radiation orchemotherapy, senescence may be triggered in a variety of other organs,leading to long-term ramifications for the patient. Long-termramifications may include reduced quality of life predisposing thesubject to disabilities and comorbidities. For example, a subject thathas received DNA-damaging therapy may experience a disproportionatedecline in physical function, such as inability to walk up stairs or toreach up to put things onto shelves and/or increased functionaldisabilities such as difficulty, eating, dressing and maintainingadequate hygiene. Additionally, late effects of ionizing radiation mayinclude long-term bone marrow injury and/or lung fibrosis. Long-termbone marrow injury can promote hypoplastic anemia and/or myelodysplasticsyndrome or leukemia. Further, the inventors demonstrated that followingionizing radiation, senescent cells in lung, muscle and brain aregreatly increased. These long-term ramifications provide a link betweenaccelerated aging and cancer treatment. A method to measure acceleratedaging may be as described in methods of measuring aging as above.Accordingly, administration of a composition comprising an inhibitor ofthe invention to a subject may prevent accelerated aging in a subjectwho has received DNA damaging therapy.

Definitions

“Alkyl” as used herein alone or as part of a group refers to saturatedmonovalent hydrocarbon radicals having straight or branched hydrocarbonchains or, in the event that at least 3 carbon atoms are present, cyclichydrocarbons or combinations thereof and contains 1 to 20 carbon atoms(C.sub.1-20alkyl), suitably 1 to 10 carbon atoms (C.sub.1-10alkyl),preferably 1 to 8 carbon atoms (C.sub.1-8alkyl), more preferably 1 to 6carbon atoms (C.sub.1-4alkyl), and even more preferably 1 to 4 carbonatoms (C.sub.1-4alkyl). Examples of alkyl radicals include methyl,ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl,pentyl, isoamyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyland the like.

“Alkenyl” as used herein alone or as part of a group refers tomonovalent hydrocarbon radicals having a straight or branchedhydrocarbon chains having one or more double bonds and containing from 2to about 18 carbon atoms, preferably from 2 to about 8 carbon atoms,more preferably from 2 to about 5 carbon atoms. Examples of suitablealkenyl radicals include ethenyl, propenyl, alkyl, 1,4-butadienyl andthe like.

“Alkynyl” as used herein alone or as part of a group refers tomonovalent hydrocarbon radicals having a straight or branchedhydrocarbon chains having one or more triple bonds and containing from 2to about 10 carbon atoms, more preferably from 2 to about 5 carbonatoms. Examples of alkynyl radicals include ethynyl, propynyl,(propargyl), butynyl and the like.

“Aryl” as used herein, alone or as part of a group, includes an organicradical derived from an aromatic hydrocarbon by removal of one hydrogen,and includes monocyclic and polycyclic radicals, such as phenyl,biphenyl, naphthyl.

“Alkoxy” as used herein, alone or as part of a group, refers to an alkylether radical wherein the term alkyl is as defined above. Examples ofalkyl ether radical include methoxy, ethoxy, n-propoxy, isopropoxy,n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like.

“Cycloalkyl” as used herein, alone or in combination, means a saturatedor partially saturated monocyclic, bicyclic or tricyclic alkyl radicalwherein each cyclic moiety contains from about 3 to about 8 carbonatoms, more preferably from about 3 to about 6 carbon atoms. Examples ofsuch cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like.

“Cycloalkylalkyl” as used herein, alone or in combination, means analkyl radical as defined above which is substituted by a cycloalkylradical as defined above. Examples of such cycloalkylalkyl radicalsinclude cyclopropylmethyl, cyclobutyl-methyl, cyclopentylmethyl,cyclohexylmethyl, 1-cyclopentylethyl, 1-cyclohexylethyl,2-cyclopentylethyl, 2-cyclohexylethyl, cyclobutylpropyl,cyclopentylpropyl, cyclohexylbutyl and the like.

“Substituted” means that one or more of the hydrogen atoms bonded tocarbon atoms in the chain or ring have been replaced with othersubstituents. Suitable substituents include monovalent hydrocarbongroups including alkyl groups such as methyl groups and monovalentheterogeneous groups including alkoxy groups such as methoxy groups.“Unsubstituted” means that the carbon chain or ring contains no othersubstituents other than carbon and hydrogen.

“Branched” means that the carbon chain is not simply a linear chain.“Unbranched” means that the carbon chain is a linear carbon chain.

“Saturated” means that the carbon chain or ring does not contain anydouble or triple bonds. “Unsaturated” means that the carbon chain orring contains at least one double bond. An unsaturated carbon chain orring may include more than one double bond.

“Hydrocarbon group” means a chain of 1 to 25 carbon atoms, suitably 1 to12 carbon atoms, more suitably 1 to 10 carbon atoms, and most suitably 1to 8 carbon atoms. Hydrocarbon groups may have a linear or branchedchain structure. Suitably the hydrocarbon groups have one branch.

“Carbocyclic group” means a saturated or unsaturated hydrocarbon ring.Carbocyclic groups are not aromatic. Carbocyclic groups are monocyclicor polycyclic. Polycyclic carbocyclic groups can be fused, spiro, orbridged ring systems. Monocyclic carbocyclic groups contain 4 to 10carbon atoms, suitably 4 to 7 carbon atoms, and more suitably 5 to 6carbon atoms in the ring. Bicyclic carbocyclic groups contain 8 to 12carbon atoms, preferably 9 to 10 carbon atoms in the rings.

“Heteroatom” means an atom other than carbon e.g., in the ring of aheterocyclic group or the chain of a heterogeneous group. Preferably,heteroatoms are selected from the group consisting of sulfur,phosphorous, nitrogen and oxygen atoms. Groups containing more than oneheteroatom may contain different heteroatoms.

“Heterocyclic group” means a saturated or unsaturated ring structurecontaining carbon atoms and 1 or more heteroatoms in the ring.Heterocyclic groups are not aromatic. Heterocyclic groups are monocyclicor polycyclic. Polycyclic heteroaromatic groups can be fused, spiro, orbridged ring systems. Monocyclic heterocyclic groups contain 4 to 10member atoms (i.e., including both carbon atoms and at least 1heteroatom), suitably 4 to 7, and more suitably 5 to 6 in the ring.Bicyclic heterocyclic groups contain 8 to 18 member atoms, suitably 9 or10 in the rings.

“Isomer”, “isomeric form”, “stereochemically isomeric forms” or“stereolsomeric forms”, as used herein, defines all possible isomeric aswell as conformational forms, made up of the same atoms bonded by thesame sequence of bonds but having different three-dimensional structureswhich are not interchangeable, which compounds or intermediates obtainedduring said process may possess. Unless otherwise mentioned orindicated, the chemical designation of a compound encompasses themixture of all possible stereochemically isomeric forms which saidcompound may possess. Said mixture may contain all diastereoisomers,epimers, enantiomers and/or conformers of the basic molecular structureof said compound. More in particular, stereogenic centers may have theR- or S-configuration, diastereoisomers may have a syn- oranti-configuration, substituents on bivalent cyclic saturated radicalsmay have either the cis- or trans-configuration and alkenyl radicals mayhave the E or Z-configuration. All stereochemically isomeric forms ofsaid compound both in pure form or in admixture with each other areintended to be embraced within the scope of the present invention.

EXAMPLES

The following examples illustrate various iterations of the invention.However, those of skill in the art should, in light of the presentdisclosure, appreciate that many changes can be made in the specificembodiments which are disclosed and still obtain a like or similarresult without departing from the spirit and scope of the invention.

Example 1 Piperlongumine (PL) Selectively Kills Senescent WI38 Cells ina Dose- and Time-Dependent Manner

Normal WI38 cells (FIG. 1A), IR-induced senescent cells (FIG. 1B) andreplicative senescent cells (FIG. 1C) were incubated with increasingconcentrations of PL. At 72 hours post-treatment cell viability wasmeasured. Concentrations of PL up to 10 μM had no significant effect onnormal WI38 cells (FIG. 1A). However, concentrations as low as 5 μM PLsignificantly reduced the number of viable cells in both IR-inducedsenescent cells and replicative senescent cells (FIG. 1B, C). IR-inducedsenescent WI38 cells were treated with 10 μM PL and viable cells weremeasured at 0, 1, 2, 3, 6, 12, 16, 24, 48 and 72 hours after treatment(FIG. 1D) or the cells were incubated with 10 μM PL for 1, 3, 6, 12, 24,48 or 72 hours and cell viability was measured at 72 h (FIG. 1E). Therewas a significant reduction in viable cells from 6 hours on in bothtreatment scenarios (FIG. 1D, E).

Example 2 PL Selectively Increases ROS Production in Senescent Cells

PL selectively targets senescent cells as measured by the LD50 value.Other ROS target drugs such as auranofin, buthionine sulfoximine, anddecyl-triphenylphosphonium have approximately equivalent LD50 valuesbetween normal and senescent cells (Table 1).

PL selectively increases ROS in senescent WI38 in a time-dependentmanner. Normal (FIG. 2A) and IR-induced senescent WI38 cells (FIG. 2B)were incubated with vehicle (Veh), 200 mM H₂O₂ for 30 min as a positivecontrol, or 10 μM PL. Reactive oxygen species (ROS) was measured atvarious times (1, 3, 6, 12, 24 hours) after PL treatment and expressedas percent of control. Data are presented as mean±SE of threeindependent assays. PL significantly increased ROS production at 24hours.

TABLE 1 LD50 value of ROS target drugs and PL against normal and varioussenescent WI38 cells. LD50 LD50 (IR- LD50 Ratio (normal) senescent)(Replicative) MOA or (normal/ (uM) (uM) (uM) Target senescent) Auranofin5.13 5.34 Nd TrxR inhibitor/ 1.0 ROS γ-GSC Buthionine 2340.19 2391.51 Ndinhibitor/ROS 1.0 sulfoximine Decyl- 1.41 0.98 Nd Mitochondria/ROS 1.4triphenylphosphonium PL 24.25 7.05 6.35 ROS 3.44-3.82 γ-GSC:γ-glutamylcysteine synthetase; TrxR: Thioredoxin reductase

Example 3 N-Acetyl-Cysteine (NAC) but not the Pan-Caspase Inhibitor QCDAttenuates PL Induced Cell Death in Senescent Cells

IR-induced senescent WI38 cells were pre-treated with the pan-caspaseinhibitor Q-VD-OPh (20 μM) (FIG. 3B) or the antioxidant NAC (1 mM) (FIG.3A) for 2 hours and then incubated with vehicle (VEH) or 10 μM PL for 72h. Cell viability was measured and expressed as percent of control. Dataare presented as mean±SE of three independent assays. NAC selectivelyinhibited PL induced cell death in senescent cells

Example 4 PL and ABT263 Can Synergistically Kill Senescent Cells

Normal WI38 cells were incubated with vehicle (VEH), 1.25 μM ABT263, PL(10 uM), or both (FIG. 4A). IR-induced senescent WI38 cells wereincubated with vehicle (VEH), 1.25 μM ABT263, PL (5 or 10 uM), or both(FIG. 4B). IR-induced senescent WI38 cells were incubated with vehicle(VEH), PL (10 uM), increasing concentrations of ABT263, or both (FIG.4C). Cell viability was measured at 72 h and expressed as percent ofcontrol. Data are presented as mean±SE of three independent assays. Thecombination of PL and ABT263 synergistically killed senescent cells.

Example 5 Piperlongumine (PL) Derivative XL-11155B (5) Selectively KillsSenescent WI38 Cells in a Dose- and Time-Dependent Manner

Normal WI38 cells (FIG. 5A) and IR-induced senescent cells (FIG. 5B)were incubated with increasing concentrations of XL-11155B. At 72 hourspost-treatment cell viability was measured. The LD₅₀ of XL-11155B onnormal WI38 cells was 4.85 μM (FIG. 5A). However, The LD₅₀ of XL-11155Bon IR-induced senescent WI38 cells was 1.72 μM (FIG. 5B).

Example 6 Evaluation of Compounds of Formula I and Formula II and TheirAbility to Selectively Kill Senescent Cells

Normal WI38 cells and IR-induced senescent cells were incubated withincreasing concentrations of compounds of Formula I and Formula II. At72 hours post-treatment cell viability was measured and LD50 wascalculated. Table 2 depicts the LD50 values of the compounds of FormulaI and compounds of Formula II against normal and IR-induced senescentcells.

TABLE 2 LD50 of compounds of Formula I and Formula II against normal andsenescent cells. LD50 (uM) non- IR- senescent senescent Ratio NameStructure cells (NSC) cells (SC) (NSC/SC) GZ-11702

169.70 208.00 0.82 GZ-11718

125.50 45.51 2.76 XL-11155A

34.18 9.10 3.76 GZ-11717B

34.25 16.11 2.13 GZ-11719-by

84.71 47.15 1.80 GZ-11719

1.62 0.90 1.80 XL-11155B

5.60 1.95 2.87 XL-11178B

12.1 7.92 1.53 XL-11178A

>40 >40 — XL-11194

10.28 4.02 2.56 GZ-11725A

22.4 6.6 3.4  GZ-11725B

15.73 5.35 2.94 GZ-11728

7.7 1.0 7.6  XL-12104

4.24 1.62 2.62 XZ-12107

23.96 4.77 5.02 GZ-11734

7.16 1.39 5.14 XZ-12111B

>40 >40 — XL-12115

52.62 30.00 1.75 XZ 12040

5.78 2.4 2.41 XZ-12045

13.93 4.38 3.18 XZ-12047

21.86 10.77 2.03 XZ 12043

23.53 15.71 1.50 XZ-12041

22.07 9.97 2.21 XZ-12052

80.00 41.04 1.95 XL-12128

1.95 0.67 2.91 XL-12124

ND ND — XZ-12208

10.16 2.66 3.82 XZ-12209

9.02 5.40 1.67 XZ-12210

7.31 3.92 1.91 XZ-12213

4.85 4.00 1.21 XZ-12215

12.35 9.478 1.30 XZ-12216

8.74 5.18 1.69 PS-12611

8.48 6.13 1.38 PS-12612

8.58 5.86 1.46 PS-12626

4.51 1.20 3.77 PS-12624

15.75 12.90 1.22 PS-12636

18.78 6.76 2.78 PS-12633

21.04 8.33 2.53 PS-12634

7.67 7.11 1.08 PS-12637

17.50 6.41 2.73 PS-12638

20.14 7.10 2.84 PS-12639

13.46 6.47 2.08 PS-12640

14.37 7.72 1.86 XZ-12237

12.67 5.32 2.38 XZ-12224

6.41 2.20 2.92 XZ-12235

12.85 7.77 1.65 XL-12137A

>40 >40 — XL-12137B

>40 >40 — XL-12138

12.41 2.09 5.93 PS-12644

24.20 18.99 1.27 PS-12643

15.41 6.74 2.28 PS-12641

3.73 5.12 0.73 PS-12646

35.34 18.57 1.90 PS-12654

22.10 17.02 1.30

Example 7 Synthesis of Exemplary Piperlongumine Derivatives of Formula(I)

General Method:

To a solution of acid (1 eq) in dry THF was added Et₃N (1.15 eq) andpivaloyl chloride (1.1 eq). The reaction mixture was stirred for 45-60min for completion (monitored by TLC). The mixture was filtered and thefiltrat was used directly for the next step.

To a stirred solution of 5,6-dihydropyridin-2(1H)-one (0.8 eq) in dryTHF was added 0.88 eq of n-BuLi under inert atmosphere at −78° C. Afterstirred for 0.5 h, the mixed anhydride solution prepared from above wasadded and the resulting reaction mixture was stirred for 1 h beforequenched with saturated NH₄Cl (0.5 mL). The mixture was extracted withethylacetate (10 mL×3). The organic phases were combined, washed withsat. NaCl, dried over anhydrous Na₂SO₄, and evaporated to dryness undervacuum. The product was purified by silica gel column chromatography.

Preparation of(E)-1-(3-(3-fluoro-4-chlorophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 28%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.61 (d, J=15.6 Hz,1H), 7.46 (d, J=15.6 Hz, 1H), 7.41-7.27 (m, 3H), 6.96 (m, 1H), 6.05 (dd,J=9.6, 2 Hz, 1H), 4.04 (t, J=6 Hz, 2H), 2.49 (m, 2H) ppm; ¹³C-NMR (100MHz, CDCl₃) δ 168.39, 165.75, 159.43, 156.95, 145.76, 140.68 (d, J=2.3Hz), 135.74 (d, J=7.6 Hz), 130.88, 125.64, 124.72 (d, J=3 Hz), 123.66,122.44 (d, J=18 Hz), 115.55 (d, J=21.3 Hz), 41.60, 24.72 ppm.

Preparation of(E)-1-(3-(3-chloro-4-fluorophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 30%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.61 (m, 2H), 7.42(m, 2H), 7.13 (t, J=8.8 Hz, 1H), 6.96 (m, 1H), 6.05 (m, 1H), 4.03 (t,J=6.8 Hz, 2H), 2.48 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 168.42,165.77, 160.10, 157.58, 145.74, 140.62 (d, J=1.6 Hz), 132.45 (d, J=4.6Hz), 130.05, 128.22 (d, J=7.5 Hz), 125.64, 122.93 (d, J=2.3 Hz), 121.59(d, J=18.2 Hz), 116.94 (d, J=11 Hz), 41.59, 24.73 ppm.

Preparation of(E)-1-(3-(3,4-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 58%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.71 (d, J=15.6 Hz,1H), 7.40 (d, J=15.2 Hz, 1H), 7.15 (dd, J=8.4, 2 Hz, 1H), 7.10 (d, J=0.8Hz, 1H), 6.92 (m, 1H), 6.84 (d, J=8.4 Hz, 1H), 6.04 (m, 1H), 4.03 (t,J=6 Hz, 2H) 3.90 (s, 6H), 2.46 (m, 2H) ppm.

Preparation of(E)-1-(3-(2,5-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 31%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.05 (d, J=15.6 Hz,1H), 7.52 (d, J=15.6 Hz, 1H), 7.11 (d, J=2.8 Hz, 1H), 6.90 (m, 2H), 6.83(d, J=8.8 Hz, 1H), 6.03 (d, J=10 Hz, 1H), 4.02 (m, 2H), 3.83 (s, 3H),3.78 (s, 3H), 2.45 (m, 2H) ppm.

Preparation of(E)-1-(3-(2,3-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 57%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.03 (d, J=15.6 Hz,1H), 7.51 (d, J=16 Hz, 1H), 7.21 (d, J=8 Hz, 1H), 7.02 (m, 1H), 6.90 (d,J=7.6 Hz, 2H), 6.00 (dd, J=9.6, 1.6 Hz, 1H), 4.01 (m, 2H), 3.85 (s, 6H),2.45 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 169.10, 165.75, 153.04,148.53, 145.46, 138.06, 129.25, 125.76, 124.05, 123.08, 119.60, 113.71,61.38, 55.82, 41.59, 24.76 ppm.

Preparation of(E)-1-(3-(2,6-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 46%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.13 (d, J=16 Hz,1H), 7.82 (d, J=16 Hz, 1H), 7.17 (m, 1H), 6.81 (m, 1H), 6.46 (dd, J=8.4,4.4 Hz, 2H), 5.94 (d, J=10 Hz, 1H), 3.94 (m, 2H), 3.80 (s, 6H), 2.37 (m,2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 170.50, 165.68, 160.09, 145.00,134.72, 130.99, 125.99, 124.16, 112.99, 103.65, 103.61, 55.79, 55.74,41.66, 24.80 ppm. GC-MS: 287.1 (M⁺).

Preparation of(E)-1-(3-(3,5-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 66%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.63 (d, J=15.6 Hz,1H), 7.43 (d, J=15.6 Hz, 1H), 6.91 (m, 1H), 6.69 (d, J=2.4 Hz, 2H), 6.45(m, 1H), 6.00 (m, 1H), 4.00 (t, J=6.4 Hz, 2H), 3.78 (s, 6H), 2.44 (m,2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 168.84, 165.75, 160.87, 145.61,143.49, 136.93, 125.67, 122.28, 106.09, 102.47, 55.38, 41.60, 24.73 ppm.GC-MS: 287.1 (M⁺).

Preparation of(E)-1-(3-(2,4-dimethoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 53%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.03 (d, J=15.6 Hz,1H), 7.51 (m, 2H), 6.89 (m, 1H), 6.48 (dd, J=8.4, 2.4 Hz, 1H), 6.42 (d,J=2.4 Hz, 1H), 6.02 (m, 1H), 4.01 (m, 2H), 3.85 (s, 3H), 3.82 (s, 3H),2.44 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 169.60, 165.78, 162.62,159.90, 145.06, 139.20, 130.32, 126.00, 119.54, 117.30, 105.15, 98.29,55.48, 55.41, 41.60 24.80 ppm.

Preparation of(E)-1-(3-(3-nitro-4-methoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 47%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.03 (d, J=2.4 Hz,1H), 7.74 (dd, J=8.4, 2.4 Hz, 1H), 7.64 (d, J=15.6 Hz, 1H), 7.44 (d,J=15.6 Hz, 1H), 7.08 (d, J=8.8 Hz, 1H), 6.96 (m, 1H), 6.04 (m, 1H), 4.03(m, 2H), 3.99 (s, 3H), 2.48 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ168.42, 165.78, 153.78, 145.74, 140.29, 139.80, 133.58, 127.88, 125.65,122.47, 113.69, 56.71, 41.63, 24.74 ppm.

Preparation of(E)-1-(3-(3-amino-4-methoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

A mixture of PS12643 (92 mg, 305 mmol), Fe (68.2 mg, 1.22 mol), andNH₄Cl (65.2 mg, 1.2 mol) in 6 mL of EtOH and 3 mL H₂O was reacted at40-50° C. for 3 h. After the completion of the reaction (monitored byTLC), the reaction was cooled to rt, neutralized with sat. NaHCO₃,extracted with dichloromethane. The organic phase was washed with brine,dried over anhydrous Na₂SO₄, and evaporated to dryness under vacuum. Theproduct was purified by silica gel column chromatography to afford thetarget compound (74 mg) as a yellow solid, yield: 90%. ¹H-NMR (400 MHz,CDCl₃) δ 7.66 (d, J=15.6 Hz, 1H), 7.36 (d, J=15.6 Hz, 1H), 6.99-6.89 (m,3H), 6.77 (d, J=10.8 Hz, 1H), 6.03 (m, 1H), 4.02 (m, 2H), 3.86 (s, 3H),2.45 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 169.16, 165.82, 149.31,145.22, 144.35, 136.28, 128.16, 125.94, 120.78, 119.09, 113.33, 110.03,55.52, 41.55, 24.79 ppm.

Preparation of(E)-1-(3-(3-acetylamino-4-methoxylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

A mixture of PS12644 (68 mg, 0.25 mmol), Ac₂O (35 μL, 0.37 mmol), andEt₃N (52 μL, 0.37 mmol) in dichloromethane was allowed stirred at rtovernight. After the completion of the reaction (monitored by TLC), thereaction was extracted with dichloromethane and water. The organic phasewas washed with brine, dried over anhydrous Na₂SO₄, and evaporated todryness under vacuum. The product was purified by silica gel columnchromatography to afford the target compound (55 mg) as a yellow solid,yield: 70%. ¹H-NMR (400 MHz, CDCl₃) δ 8.60 (s, 1H), 7.70 (m, 2H), 7.36(d, J=15.6 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 6.91 (m, 1H), 6.84 (d, J=8.8Hz, 1H), 6.02 (d, J=10 Hz, 1H), 4.01 (s, 2H), 3.90 (s, 3H), 2.45 (m,2H), 2.20 (s, 3H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 169.14, 168.11,165.69, 149.12, 145.17, 143.52, 128.35, 127.89, 125.88, 124.04, 120.39,119.84, 109.88, 55.83, 41.63, 24.88, 24.80 ppm. GC-MS: 313.1 (M⁺).

Preparation of(E)-1-(3-(4-dimethylaminophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 52%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.75 (d, J=15.6 Hz,1H), 7.48 (d, J=8.4 Hz, 1H), 7.36 (d, J=16 Hz, 1H), 6.89 (m, 1H), 6.65(d, J=8.8 Hz, 2H), 6.03 (m, 1H), 4.02 (t, J=6.4 Hz, 2H), 3.01 (s, 6H),2.44 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 169.45, 165.84, 151.70,144.96, 144.94, 130.14, 126.09, 122.96, 116.29, 111.71, 41.57, 40.12,24.82 ppm.

Preparation of(E)-1-(3-(1-acetyl-1H-indol-3-yl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 25%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.45 (d, J=8 Hz,1H), 7.93-7.86 (m, 2H), 7.75-7.69 (m, 2H), 7.38 (m, 2H), 6.95 (m, 1H),6.06 (d, J=9.6 Hz, 1H), 4.07 (m, 2H), 2.65 (s, 3H), 2.48 (m, 2H) ppm;¹³C-NMR (100 MHz, CDCl₃) δ 168.85, 168.40, 165.86, 145.50, 136.57,134.96, 127.97, 127.60, 126.00, 125.87, 124.57, 121.96, 120.37, 119.13,116.75, 41.56, 24.77, 23.98 ppm.

Preparation of(E)-1-(3-(pyridine-2-yl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 44%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 8.61 (d, J=4 Hz,1H), 7.80 (d, J=15.2 Hz, 1H), 7.68-7.64 (m, 2H), 7.45 (d, J=8 Hz, 1H),7.20 (dd, J=7.6, 4.8 Hz, 1H), 6.92 (m, 1H), 6.01 (d, J=9.6 Hz, 1H), 4.00(m, 2H), 2.45 (m, 2H) ppm. GC-MS: 228.1 (M⁺).

Preparation of(E)-1-(3-(pyridine-3-yl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 61%; yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.77 (d, J=2.0 Hz,1H), 8.58 (dd, J=4.8, 1.6 Hz, 1H), 7.92-7.89 (m, 1H), 7.70 (d, J=15.6Hz, 1H), 7.56 (d, J=15.6 Hz, 1H), 7.34-7.30 (m, 1H), 6.99-6.95 (m, 1H),6.07-6.03 (m, 1H), 4.05 (t, J=6.4 Hz, 2H), 2.52-2.47 (m, 2H) ppm; ¹³CNMR (100 MHz, CDCl₃) δ 168.3, 165.7, 150.6, 149.9, 145.8, 139.3, 134.2,130.8, 125.6, 124.0, 123.6, 41.6, 24.7 ppm

Preparation of(E)-1-(3-(furan-3-yl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 62%; ¹H NMR (400 MHz, CDCl₃) δ 7.53-7.35 (m, 3H), 6.95-6.91 (m,1H), 6.63 (d, J=3.2 Hz, 1H), 6.47-6.45 (m, 1H), 6.05-6.01 (m, 1H), 4.02(t, J=6.4 Hz, 2H), 2.49-2.43 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ168.7, 165.6, 151.6, 145.4, 144.5, 130.1, 125.7, 119.4, 114.7, 112.2,41.6, 24.7 ppm; GC-MS 217 (M⁺)

Preparation of(E)-1-(3-(2-(trifluoromethyl)phenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 21%; yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.08-8.03 (m, 1H),7.79 (d, J=7.6 Hz, 1H), 7.67 (d, J=7.6 Hz, 1H), 7.56-7.40 (m, 3H),6.97-6.92 (m, 1H), 6.05-6.01 (m, 1H), 4.04 (t, J=6.4 Hz, 2H), 2.50-2.45(m, 2H) ppm.

Preparation of(E)-2-(6-oxo-1,2,3,6-tetrahydropyridine-1-carbonyl)-3-(3,4,5-trimethoxyphenyl)acrylonitrile

Yield 19%; ¹H NMR (400 MHz, CDCl₃) δ 7.75 (s, 1H), 7.19 (s, 2H),6.97-6.91 (m, 1H), 6.04-6.00 (m, 1H), 3.89-3.84 (m, 11H), 2.55-2.50 (m,2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ 167.5, 165.2, 153.2, 151.9, 146.4,142.2, 127.2, 124.6, 116.2, 108.2, 107.6, 61.1, 56.3, 43.6, 24.7 ppm;GC-MS 342 (M⁺).

Preparation of (E)-1-(3-(4-bromophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 58%; yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.65 (d, J=15.6 Hz,1H), 7.52-7.40 (m, 5H), 6.96-6.90 (m, 1H), 6.05-6.01 (m, 1H), 4.03 (t,J=6.4 Hz, 2H), 2.50-2.43 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ 168.9,165.8, 145.6, 142.0, 134.0, 132.0, 130.0, 125.7, 124.2, 122.5, 41.6,24.8 ppm

Preparation of(E)-1-(3-(4-chlorophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 74%; ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J=16.0 Hz, 1H), 7.51-7.45(m, 3H), 7.35-7.30 (m, 2H), 6.97-6.91 (m, 1H), 6.06-6.01 (m, 1H), 4.03(t, J=6.4 Hz, 2H), 2.50-2.43 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ168.7, 165.8, 145.6, 142.0, 135.8, 133.6, 129.4, 129.0, 125.7, 122.4,41.6, 24.8 ppm

Preparation of(E)-1-(3-(2,4-dichlorophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 56%; yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.03 (d, J=15.6 Hz,1H), 7.64 (d, J=8.8 Hz, 1H), 7.49-7.42 (m, 2H), 7.27-7.25 (m, 1H),6.99-6.94 (m, 1H), 6.04 (d, J=10.0 Hz, 1H), 4.05 (t, J=6.4 Hz, 2H),2.52-2.47 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ 168.3, 165.8, 145.8,137.7, 135.9, 135.6, 132.0, 129.8, 128.7, 127.4, 125.6, 124.8, 41.6,24.7 ppm

Preparation of(E)-1-(3-(4-fluoro-3-methylphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 63%; ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J=16.0 Hz, 1H), 7.44-7.35(m, 3H), 7.01-6.90 (m, 2H), 6.05-6.02 (m, 1H), 4.03 (t, J=6.4 Hz, 2H),2.50-2.45 (m, 2H), 2.27 (s, 3H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ 168.8,165.8, 162.4 (d, J=248 Hz), 145.5, 142.7, 131.3 (d, J=6 Hz), 131.0 (d,J=4 Hz), 127.8, 127.7, 125.8, 121.2 (d, J=2 Hz), 115.4 (d, J=23 Hz),41.6, 24.8, 14.4 (d, J=4 Hz) ppm

Preparation of(E)-1-(3-(3-fluoro-4-methoxyphenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 41%; ¹H NMR (400 MHz, CDCl₃) δ 7.65 (d, J=15.6 Hz, 1H), 7.40-7.30(m, 3H), 7.00-6.92 (m, 2H), 6.05-6.02 (m, 1H), 4.04 (t, J=6.4 Hz, 2H),3.92 (s, 3H), 2.50-2.46 (m, 2H) ppm.

Preparation of(E)-1-(3-(4-nitrophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield 55%; yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.25-8.20 (m, 2H),7.75-7.70 (m, 3H), 7.59 (d, J=16.0 Hz, 1H), 7.02-6.95 (m, 1H), 6.08-6.04(m, 1H), 4.06 (t, J=6.4 Hz, 2H), 2.55-2.49 (m, 2H) ppm; ¹³C NMR (100MHz, CDCl₃) δ 168.1, 165.8, 148.2, 146.0, 141.3, 139.9, 128.8, 126.1,125.5, 124.0, 41.6, 24.7 ppm; GC-MS 272 (M⁺).

Preparation of(E)-1-(3-(4-aminophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

XZ12224 (200 mg, 0.74 mmol), iron powder (206 mg, 3.68 mmol), and NH₄Cl(197 mg, 3.68 mmol) in a mixture of EtOH (15 mL) and water (3 mL) wereheated to 55° C. for 4 h. The mixture was then cooled to rt andextracted with EtOAc. The organic layer was separated washed with brineand dried over anhydrous Na₂SO₄. The combined organic layers wereevaporated to dryness to give crude product, which was purified by flashcolumn chromatography on silica gel using hexane/EtOAc (1:1) as eluent.Yield 57%; ¹H NMR (400 MHz, CDCl₃) δ 7.69 (d, J=15.2 Hz, 1H), 7.40-7.30(m, 3H), 6.92-6.85 (m, 1H), 6.61 (d, J=8.2 Hz, 2H), 6.01 (d, J=9.6 Hz,1H), 4.02-3.97 (m, 4H), 2.45-2.40 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃)δ 169.4, 165.9, 148.8, 145.2, 144.5, 130.2, 126.0, 125.2, 117.3, 114.7,41.6, 24.8 ppm; GC-MS 242 (M⁺).

Preparation of(E)-1-(3-(4-acetylaminophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

XZ12235 (36 mg, 0.15 mmol), Ac₂O (22 μL, 0.23 mmol) and trimethylamine(32 μL, 0.23 mmol) in 5 mL DCM were stirred at room temperature for 16h. The mixture was concentrated and purified by flash columnchromatography on silica gel using hexane/EtOAc (1:1) as eluent. Yield80%; ¹H NMR (400 MHz, CDCl₃) δ 7.69-7.38 (m, 7H), 6.94-6.91 (m, 1H),6.02 (d, J=9.6 Hz, 1H), 4.01 (t, J=6.0 Hz, 2H), 2.47-2.44 (m, 2H), 2.15(s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 169.0, 168.4, 165.9, 144.6, 143.1,139.7, 130.8, 129.2, 125.8, 120.6, 119.6, 41.6, 24.8, 24.7 ppm

Preparation of(E)-1-(3-(2-fluorophenyl)prop-2-enoyl)-5,6-dihydropyridin-2(1H)-one

Yield: 48%; yellow solid. ¹H-NMR (400 MHz, CDCl₃) δ 7.87 (d, J=16 Hz,1H), 7.60 (m, 1H), 7.54 (d, J=15.6 Hz, 1H), 7.34 (m, 1H), 7.15-7.05 (m,2H), 6.94 (m, 1H), 6.03 (d, J=9.6 Hz, 1H), 4.03 (m, 2H), 2.48 (m, 2H)ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 168.79, 165.73, 162.60, 160.08, 145.57,135.63 (d, 3.1 Hz), 131.38 (d, 8.4 Hz), 128.92 (d, 3 Hz), 125.71, 124.10(m), 123.19, 116.10 (d, 22 Hz), 41.61, 27.02 ppm. GC-MS: 245.1 (M⁺).

Preparation of1-(3-(3,4,5-trimethoxyphenyl)propioloyl)-5,6-dihydropyridin-2(1H)-one

Yield 51%; ¹H NMR (400 MHz, CDCl₃) δ 6.99-6.94 (m, 1H), 6.89 (s, 2H),6.09-6.05 (m, 1H), 4.06 (t, J=6.4 Hz, 2H), 3.88 (s, 3H), 3.86 (s, 6H),2.53-2.47 (m, 2H) ppm; ¹³C NMR (100 MHz, CDCl₃) δ 163.7, 153.1, 153.0,145.6, 140.8, 125.1, 115.2, 110.4, 93.9, 83.1, 61.0, 56.2, 40.8, 24.4ppm

Example 8 Synthesis of Exemplary Piperlongumine Derivatives of Formula(II) Preparation of(E)-3-methylene-1-(3-(3,4,5-trimethoxyphenyl)acryloyl)piperidine-2-one

3-(Hydroxymethyl)piperidin-2-one

A mixture of ethyl 2-oxopiperidine-3-carboxylate (1 g, 5.84 mmol) andcalcium chloride (0.86 g, 5.85 mmol) in ethanol (10 mL) was cooled to 0°C. and treated with sodium borohydride (0.22 g, 5.84 mmol). Theresulting mixture was allowed to warm to room temperature and stirredfor 6 hrs. The reaction was then quenched with acetone and the pH of themixture was adjusted to pH to 3-4 with 1.0 M HCl. The resulting mixturewas extracted with methylene chloride, organic phases were combined,dried over Na₂SO₄, filtered, and evaporated to dryness under vacuum. Theproduct was purified by silica gel column chromatography (377 mg, 50%yield). ¹H-NMR (400 MHz, CDCl₃) δ 6.65 (br, 1H), 3.90 (br, 1H), 3.69 (m,2H), 3.27 (m, 2H), 2.48 (m, 1H), 1.88 (m, 2H), 1.73 (m, 1H), 1.50 (m,1H) ppm.

3-Methylene-2-piperidinone

Cuprous iodine (14 mg, 0.0074 mmol) was added to a stirred solution of3-(Hydroxymethyl)piperidin-2-one (93 mg, 0.72 mmol) anddicyclohexylcarbodiimide (DCC) (186 mg, 0.9 mmol) in toluene (3 mL). Theresulting mixture was stirred under reflux for 1.5 hrs and cooled toroom temperature. Water was added and the mixture stirred overnight.Ethylacetate was added and the mixture was filtered. The aqueous phasewas extracted with ethyl acetate and the combined organic phases werewashed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The product was purified by silica gel columnchromatography (50 mg, 63% yield). ¹H-NMR (400 MHz, CDCl₃) δ 7.05 (br,1H), 6.18 (s, 1H), 5.29 (s, 1H), 3.37 (t, J=6 Hz, 2H), 2.56 (t, J=6 Hz,2H), 1.84 (m, 2H) ppm.

(E)-3-methylene-1-(3-(3,4,5-trimethoxyphenyl)acryloyl) 2-piperidinone

A solution of 3-Methylene-2-piperidinone (50 mg, 0.45 mmol) in THF (3mL) was cooled to −78° C., n-BuLi (2.5M in hexane) (0.18 mL, 0.45 mmol)was added slowly. The resulting mixture was allowed to stir at −78° C.for 1 hr. A solution of (E)-3-(3,4,5-trimethoxyphenyl)acryloyl chloride(115 mg, 0.45 mmol) in THF was then added dropwise. After addition, themixture was stirred under −78° C. for 1 hr. The reaction was thenquenched with aqueous ammonium chloride solution and was extracted withethylacetate. The combined organic phases was washed with brine, driedover Na₂SO₄, filtered, and evaporated to dryness under vacuum. Theproduct was purified by silica gel column chromatography (53 mg, 36%yield). ¹H-NMR (400 MHz, CDCl₃) δ 7.63 (d, J=15.2 Hz, 1H), 7.45 (d,J=15.2 Hz, 1H), 6.79 (s, 2H), 6.39 (d, J=1.6 Hz, 1H), 5.51 (dd, J=3.6,1.6 Hz, 1H), 3.88 (s, 6H), 3.86 (s, 2H), 3.85 (m, 2H), 2.64 (m, 2H),1.93 (m, 2H) ppm; ¹³C-NMR (100 MHz, CDCl₃) δ 170.06, 166.97, 153.45,143.66, 140.06, 138.78, 130.84, 125.86, 121.59, 105.61, 61.06, 56.31,45.15, 29.36, 22.54 ppm.

Preparation of(E)-3-methylene-1-(3-(3,4,5-trimethoxyphenyl)acryloyl)pyrrolidin-2-one

3-(Hydroxymethyl)pyrrolidin-2-one

Ethyl 2-oxopyrrolidine-3-carboxylate (250 mg, 1.59 mmol) was added to asuspension of NaBH₄ (151 mg, 3.98 mmol) and CaCl₂ (585 mg, 3.98 mmol) inethanol (5 mL) under 0° C., the resulting mixture was stirred overnightat rt. The reaction was quenched with 1M HCl solution, then extractedwith CH₂Cl₂/MeOH(10/1) for 10 to 20 times, the combined organic phaseswere dried over Na₂SO₄, filtered, and evaporated to dryness undervacuum. The product was purified by silica gel column chromatography (90mg, 50% yield). ¹H NMR (400 MHz, CDCl₃): 6.053 (br. 1H), 3.881 (m, 1H),3.736 (m, 1H), 3.384 (m, 2H), 2.626 (m, 1H), 2.247 (m, 1H), 1.954 (m,1H) ppm.

3-Methylenepyrrolidin-2-one

3-(Hydroxymethyl)pyrrolidin-2-one (16 mg, 0.139 mmol) was treated withCuI (2.6 mg, 0.0139 mmol) and DCC (36 mg, 0.174 mmol) in toluene underreflux for 3 h. The reaction mixture was cooled to rt and quenched withseveral drops of water, followed by filtration, the filtrate wasconcentrated and purified through column to obtain 11 mg of product (82%yield). ¹H NMR (400 MHz, CDCl₃): 6.2 (br. 1H), 6.011 (s, 1H), 5.382 (s,1H), 3.444 (t, J=6.4, 2H), 2.911 (m, 2H) ppm.

(E)-3-methylene-1-(3-(3,4,5-trimethoxyphenyl)acryloyl)pyrrolidin-2-one

3-Methylenepyrrolidin-2-one (14 mg, 0.144 mmol) was dissolved in THF andcooled to −78° C., 1.6 M n-BuLi (90 μL) was added dropwise under N₂protection, the resulting mixture was stirred under −78° C. for 1 h.(E)-t-butyl 3-(3,4,5-trimethoxyphenyl)acrylate (1.5 eq) was added. Afterstirring at −78° C. for 1 h, the reaction was quenched with NH₄Clsolution, extracted with EtOAc for 3 times, the combined organic phaseswere washed with brine and dried over Na₂SO₄, filtered, and evaporatedto dryness under vacuum. The residue was purified by flash columnchromatography to afford 22 mg product (49% yield). ¹H NMR (400 MHz,CDCl₃): δ 7.970 (d, J=15.6, 1H), 7.790 (d, J=15.6, 1H), 6.854 (s, 2H),6.269 (t, J=2.8, 1H), 5.590 (t, J=2, 1H), 3.903 (s, 6H), 3.891 (s, 3H),3.885-3.852 (m, 2H); ¹³CNMR (100 MHz, CDCl₃) δ 167.903, 167.015,153.537, 145.932, 140.461, 140.225, 130.564, 120.979, 118.346, 105.832,61.132, 56.336, 42.554, 23.073 ppm.

General Procedures for the Following Compounds

To a solution of(E)-1-(3-(3,4,5-trimethoxyphenyl)acryloyl)piperidin-2-one (150 mg, 0.47mmol) in THF at −78° C. was added LHMDS (1.0 M, 0.56 mL, 0.564 mmol).The resulting solution was stirred at this temperature for 1 h. Aromaticaldehyde (1 eq; dissolved in small amount of THF) was added dropwise.After stirring at −78° C. for an additional hour, ammonium chloridesolution was added to quench the reaction, the mixture was extractedwith EtOAc for 3 times. The combined organic phases were washed withwater and brine, dried over Na₂SO₄, filtered, and evaporated to drynessunder vacuum. The residue was purified by flash column chromatography toafford the corresponding hydroxyl intermediate (30%-50% yield).

Methanesulfonyl chloride (1.2 eq) was added to a solution of the aboveintermediate and TEA (1.5 eq) in toluene under ice bath. After stirringat rt for 1 h, additional 2.5 eq TEA was added. The resulting mixturewas heated to reflux for 5.5 h. Upon cooled to rt, the reaction wasquenched with water and extracted with ethylacetate. The combinedorganic fractions were washed with brine, dried over Na₂SO₄, filtered,and evaporated to dryness under vacuum. The residue was purified byflash column chromatography to afford the desired product (75%-90%yield).

XL-12104: ^(1H) NMR (400 MHz, CDCl₃): δ 8.038 (s, 1H), 7.678 (d, 1H,J=15.6), 7.551 (d, 1H, J=15.6), 7.453 (m, 1H), 7.311-7.269 (m, 3H),6.835 (s, 2H), 3.911 (s, 6H), 3.886 (s, 3H), 3.886-3.911 (m, 2H), 2.710(m, 2H), 1.929 (m, 2H) ppm. ¹³CNMR (100 MHz, CDCl₃) δ 170.058, 167.463,153.476, 143.716, 140.081, 136.256, 134.799, 134.078, 132.818, 130.891,130.413, 129.972, 129.950, 126.565, 121.662, 105.649, 61.086, 56.343,44.595, 26.078, 22.420 ppm.

XL-12111B: ¹H NMR (400 MHz, CDCl₃): δ 7.859 (s, 1H), 7.677 (d, J=15.2,1H), 7.509 (d, J=15.2, 1H), 7.418 (s, 1H), 7.331 (m, 3H), 6.826 (s, 2H),3.809-3.920 (m, 11H), 2.850 (m, 2H), 1.958 (m, 2H) ppm; ¹³CNMR (100 MHz,CDCl₃) δ 169.944, 167.690, 153.514, 143.807, 140.142, 137.789, 137.319,134.632, 132.143, 130.883, 129.935, 129.859, 129.024, 128.379, 121.571,105.665, 61.124, 56.313, 44.292, 26.465, 22.519 ppm.

XL-12115: ¹H NMR (400 MHz, CDCl₃): δ 8.712 (m, 1H), 7.829 (t, J=2, 1H),7.726 (td, J=2 and 8, 1H), 7.673 (d, J=15.6, 1H), 7.515 (d, J=15.6, 1H),7.423 (d, J=7.6, 1H), 7.209-7.240 (m, 1H), 6.831 (s, 2H), 3.888-3.924(m, 11H), 3.275 (m, 2H), 1.960 (m, 2H) ppm; ¹³CNMR (100 MHz, CDCl₃) δ169.952, 168.184, 155.047, 153.461, 149.651, 143.557, 140.036, 136.431,136.249, 134.928, 130.914, 127.202, 122.899, 121.693, 105.589, 61.071,56.313, 44.345, 26.647, 22.344 ppm.

Preparation of(E)-3-(2-methylpropylidene)-1-((E)-3-(3,4,5-trimethoxyphenyl)acryloyl)piperidine-2-oneand(Z)-3-(2-methylpropylidene)-1-((E)-3-(3,4,5-trimethoxyphenyl)acryloyl)piperidine-2-one

Reagents and conditions: a. LHDMS, isobutyraldehyde, THF, −78⁰C; b. 1).MsCl, TEA, THF, −20⁰C; 2). TEA, Tol., reflux. c. TFA, CH₂Cl₂, 0⁰C; d.n-BuLi, THF, −78⁰C.

tert-Butyl 3-(1-hydroxy-2-methylpropyl)-2-oxopiperidine-1-carboxylate

To a solution of tert-butyl 2-oxopiperidine-1-carboxylate (800 mg, 4mmol) in THF at −78° C. was added LHDMS (1.6 M, 2.76 mL, 4.4 mmol) underN₂ protection. After stirring at this temperature for 1 h,isobutyraldehyde (0.44 mL, 4.8 mmol) was added dropwise, the resultingmixture was allowed to stir for an additional hour before quenched byslowly adding saturated NH₄Cl solution. Diluted with EtOAc and theorganic fraction was washed with brine, dried over Na₂SO₄, filtered, andevaporated to dryness under vacuum. The residue was purified by flashcolumn chromatography to afford the desired product.

(Z/E)-tert-Butyl 3-(2-methylpropylidene)-2-oxopiperidine-1-carboxylate

To a solution of t-butyl3-(1-hydroxy-2-methylpropyl)-2-oxopiperidine-1-carboxylate (470 mg, 1.73mmol) and TEA (0.725 mL, 5.19 mmol) in toluene was added methanesulfonylchloride (0.2 mL, 2.60 mmol) at 0° C., after stirring at rt for 1 h,additional 0.725 mL TEA was then added, the resulting mixture was heatedto reflux and stirred under reflux for 6 h. Upon cooled down to rt, thereaction was quenched with water and extracted with EtOAc. The combinedorganic fractions were washed with brine, dried over Na₂SO₄, filtered,and evaporated to dryness under vacuum. The residue was purified byflash column chromatography to afford both the E isomer and Z isomerproduct with Z/E ratio at about 1 to 5.2 (25 mg Z isomer and 130 mg Eisomer, 25% overall yield for 3 steps).

E Isomer:

¹H NMR (400 MHz, CDCl₃): δ 6.718 (dt, J=10.0, 2.0 Hz, 1H), 3.624 (m,2H), 2.497 (m, 1H), 2.421 (td, J=1.6, 6.4 Hz, 2H), 1.799 (m, 2H), 1.590(s, 9H), 0.960 (s, 3H), 0.943 (s, 3H); ¹³CNMR (100 MHz, CDCl₃) δ165.679, 153.316, 149.059, 127.703, 82.677, 45.863, 28.043, 27.968,27.839, 21.737, 21.684.

Z Isomer:

¹H NMR (400 MHz, CDCl₃): δ 5.631 (dt, J=1.6, 9.6 Hz, 1H), 3.625 (t,J=6), 3.398 (m, 1H), 2.413 (m, 2H), 1.842 (m, 2H), 1.513 (s, 9H), 0.994(s, 3H), 0.977 (s, 3H); ¹³CNMR (100 MHz, CDCl₃) δ 166.423, 153.028,151.078, 127.658, 82.715, 45.597, 30.624, 28.210, 28.066, 22.966,22.830.

(Z or E)-3-(2-methylpropylidene)piperidin-2-one

To a solution of t-butyl3-(2-methylpropylidene)-2-oxopiperidine-1-carboxylate (Z or E isomer,130 mg, 0.514 mmol) in CH₂Cl₂ (8 mL) at 0° C. was added TFA (0.79 mL,10.3 mmol), the resulting mixture was allowed to stir at 0° C. for 2 h.The reaction was then quenched with saturated sodium bicarbonate underice bath and extracted with CH₂Cl₂, the combined organic fractions werewashed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The residue was purified by flash columnchromatography to afford the corresponding Z or E isomer in 90% yield.

E Isomer:

¹H NMR (400 MHz, CDCl₃): δ 7.15 (br, 1H), 6.639 (d, J=10 Hz, 1H), 3.319(m, 2H), 2.576 (m, 1H), 2.464 (m, 2H), 1.806 (m, 2H), 0.999 (s, 3H),0.982 (s, 3H); ¹³CNMR (100 MHz, CDCl₃) δ 167.334, 145.500, 126.481,42.007, 27.064, 24.385, 22.921, 22.078.

Z Isomer:

¹H NMR (400 MHz, CDCl₃): δ 5.664 (br, 1H), 5.671 (d, 10.4 Hz, 1H), 3.725(m, 1H), 3.304 (m, 2H), 2.411 (m, 2H), 1.833 (m, 2H), 0.986 (s, 3H),0.970 (s, 3H); ¹³CNMR (100 MHz, CDCl₃) δ 167.030, 149.932, 125.654,42.508, 32.400, 27.626, 23.611, 22.959.

(Z orE)-3-(2-methylpropylidene)-1-((E)-3-(3,4,5-trimethoxyphenyl)acryloyl)piperidin-2-one

To a solution of 3-(2-methylpropylidene) piperidin-2-one (Z or E isomer,10 mg, 0.065 mmol) in THF was added n-BuLi (1.6 M, 49 μL, 0.078 mmol) at−78° C. under N₂ protection, after stirring at −78° C. for 1 h,(E)-tert-butyl 3-(3,4,5-trimethoxyphenyl)acrylate (28 mg, 0.098 mmol)was added. The resulting mixture was stirred at −78° C. for 45 min andthen quenched with saturated NH₄Cl water solution. The whole mixture wasextracted with EA for several times, the combined organic phase waswashed with brine, dried over Na₂SO₄, filtered, and evaporated todryness under vacuum. The residue was purified by flash columnchromatography to afford the corresponding Z or E isomer (48% yield forZ isomer and 58% yield for E isomer).

E Isomer:

¹H NMR (400 MHz, CDCl₃): δ 7.631 (d, J=16 Hz, 1H), 7.500 (d, J=16 Hz,1H), 6.767 (d, J=11.2 Hz, 1H), 6.804 (s, 2H), 3.895 (s, 6H), 3.885 (s,3H), 3.845 (m, 2H), 2.630 (m, 1H), 2.557 (m, 2H), 1.915 (m, 2H), 1.075(s, 3H), 1.058 (s, 3H); ¹³CNMR (100 MHz, CDCl₃) δ 170.058, 168.024,153.476, 150.425, 143.367, 139.990, 131.020, 127.977, 121.920, 105.604,61.109, 56.351, 44.223, 27.960, 24.294, 22.382, 21.873.

Z Isomer:

¹H NMR (400 MHz, CDCl₃): δ 7.640 (d, J=16 Hz, 1H), 7.388 (d, 16 Hz, 1H),6.802 (s, 2H), 5.778 (d, J=9.6 Hz, 1H), 3.899 (s, 6H), 3.879 (s, 3H),3.800 (t, J=6.4 Hz, 2H), 3.418 (m, 1H), 2.504 (t, J=5.6 Hz, 2H), 1.912(q, J=6.4, 5.6 Hz, 2H), 1.047 (s, 3H), 1.030 (s, 3H); ¹³CNMR (100 MHz,CDCl₃) δ 169.625, 168.768, 153.461, 152.300, 143.314, 140.028, 130.967,127.627, 121.670, 105.680, 61.117, 56.351, 43.533, 30.373, 28.271,22.830, 22.617.

Preparation of (E)-3-methylene-1-(styrylsulfonyl)piperidine-2-one

To a solution of 3-methylenepiperidin-2-one (80 mg, 9.0 mmol) in THF at−78° C. was added n-BuLi (1.6 M, 0.54 mL, 10.8 mmol), after stirring atthis temperature for 1 h, (E)-2-phenylethenesulfonyl chloride (146 mg,9.0 mmol, dissolved in small amount of THF) was added. The resultingmixture was stirred for another 1 h and then quenched with saturatedNH₄Cl water solution. The whole mixture was extracted with EA forseveral times, the combined organic phase was washed with brine, driedover Na₂SO₄, filtered, and evaporated to dryness under vacuum. Theresidue was purified by flash column chromatography to afford thedesired product (60 mg, 30% yield).

¹H NMR (400 MHz, CDCl₃): δ 7.663 (d, J=16 Hz, 1H), 7.529 (m, 2H), 7.415(m, 3H), 7.320 (d, J=16 Hz, 1H), 6.365 (d, J=0.8 Hz, 1H), 5.483 (d, 0.8Hz, 1H), 3.891 (t, J=6 Hz, 2H), 2.598 (t, J=6 Hz, 2H), 1.969 (q, J=6 Hz,6 Hz, 2H) ppm; ¹³CNMR (100 MHz, CDCl₃) δ 164.693, 144.414, 137.182,132.317, 131.521, 129.214, 128.887, 126.148, 124.918, 46.796, 29.258,23.293 ppm.

Preparation of (E)-3-methylene-1-(3-(3, 4,5-trimethoxyphenyl)acryloyl)azepan-2-one

tert-butyl 2-oxoazepane-1-carboxylate

To a solution of ε-Caprolactam (3 g, 26.5 mmol), DMAP (647 mg, 5.3 mmol)and Et3N (10 ml, 66.3 mmol) in CH2Cl2 at 00 C was added Boc2O (8.7 g,39.75 mmol), the resulting mixture was stirred at r.t overnight.Evaporation of solvent gave a residue which was purified through columnto get 5 g product (89% yield).

¹H NMR (400 MHz, CDCl₃): δ 3.760 (t, J=2 Hz, 2H), 2.648 (m, 2H), 1.745(m, 6H), 1.522 (s, 9H). ¹³CNMR (100 MHz, CDCl₃) δ 175.826, 153.013,82.821, 46.227, 39.594, 29.318, 28.757, 28.127, 23.604.

1-tert-butyl 3-methyl 2-oxoazepane-1,3-dicarboxylate

To a solution of tert-butyl 2-oxoazepane-1-carboxylate (2.45 g, 11.5mmol) in THF at −780 C was added 1M LHMDS (13 ml, 13.2 mmol), theresulting mixture was stirred at this temperature for 45 min and methylcarbonochloridate (1 ml, 12.6 mmol) was then added, the whole mixturewas stirred for additional 1 h and quenched with quenched with saturatedNH4Cl water solution, followed by extraction with EA for several times,the combined organic phase was washed with brine and dried overanhydrous sodium sulfate. Evaporation of solvent gave a residue whichwas purified through column to get 2.7 g product. (87% yield).

¹H NMR (400 MHz, CDCl₃): δ 5.362 (br, 1H), 3.813 (s, 3H), 3.580 (m, 2H),2.116 (m, 2H), 1.746 (m, 2H), 1.550 (m, 2H), 1.458 (s, 9H); ¹³CNMR (100MHz, CDCl₃) δ 154.068, 153.150, 146.001, 111.341, 81.046, 55.099,29.280, 28.226, 28.081, 24.515, 24.014.

tert-butyl 3-methylene-2-oxoazepane-1-carboxylate

To a suspension of 1-tert-butyl 3-methyl 2-oxoazepane-1,3-dicarboxylate(90 mg, 0.33 mmol), 18-crown-6 (26 mg, 0.1 mmol), K₂CO₃ (137 mg, 1 mmol)in Toluene was added paraformaldehyde (348 mg, 11.55 mmol), theresulting mixture was stirred under reflux for 4-5 hours and then cooledto r.t. and water was added, the whole mixture was extracted with EA for3 times. The combined organic phase was washed with brine and dried overanhydrous sodium sulfate. Evaporation of solvent gave a residue whichwas purified through column to give 30 mg product. (41% yield)

¹H NMR (400 MHz, CDCl₃): δ 5.764 (s, 1H), 5.388 (s, 1H), 3.670 (m, 2H),2.424 (m, 2H), 1.740 (m, 4H), 1.527 (s, 9H); ¹³CNMR (100 MHz, CDCl₃) δ172.653, 152.846, 147.298, 123.264, 82.738, 46.060, 32.589, 29.227,28.210, 28.165.

3-methyleneazepan-2-one

To a solution of tert-butyl 3-methylene-2-oxoazepane-1-carboxylate (30mg, 0.133 mmol) in CH₂Cl₂ (2 ml) at 0° C. was added TFA (0.2 ml, 2.66mmol), the resulting solution was stirred at this temperature for 2 h.Saturated sodium bicarbonate solution was added until PH larger than 7,the mixture was then extracted with CH₂Cl₂ for 5-10 times and thecombined organic phase was washed with brine and dried over anhydroussodium sulfate. Evaporation of solvent gave a residue which was purifiedthrough column to get product (11 mg, 66% yield).

¹H NMR (400 MHz, CDCl₃): δ 6.616 (br, 1H), 5.595 (s, 1H), 5.283 (s, 1H),3.185 (m, 2H), 2.376 (m, 2H), 1.757 (m, 4H); ¹³CNMR (100 MHz, CDCl₃) δ176.053, 146.441, 120.380, 42.827, 32.771, 29.698, 28.984.

(E)-3-methylene-1-(3-(3, 4, 5-trimethoxyphenyl)acryloyl)azepan-2-one

To a solution of 3-methyleneazepan-2-one (16 mg, 0.128 mmol) in THF at−78° C. was added 1.6 M n-BuLi (88 ul) after stirring at thistemperature for 1 h, (E)-tert-butyl 3-(3,4,5-trimethoxyphenyl)acrylate(45 mg, 0.154 mmol) was then added. The resulting mixture was stirred at−78° C. for additional 1 h and then quenched with saturated NH₄Clsolution. The whole mixture was then extracted with EA for several timesand the combined organic phase was washed with brine and dried overanhydrous sodium sulfate. Evaporation of solvent gave a residue whichwas purified chromatographically to get pure product (20 mg, 45% yield).

¹H NMR (400 MHz, CDCl₃): δ 7.703 (d, J=15.2 Hz, 1H), 7.288 (d, 15.6 Hz,1H), 6.692 (s, 2H), 5.848 (d, J=1.2 Hz, 1H), 5.498 (d, J=1.2 Hz, 1H),3.893 (m, 11H), 2.495 (m, 2H), 1.807-1.852 (m, 2H), 1.200-1.781 (m, 2H);¹³CNMR (100 MHz, CDCl₃) δ 174.953, 167.690, 153.506, 147.139, 144.346,140.111, 130.716, 123.916, 119.849, 105.566, 61.109, 56.336, 44.178,32.825, 29.432, 28.119.

Preparation of(E)-3-(2-chlorobenzylidene)-1-((E)-3-(3,4,5-trimethoxyphenyl)acryloyl)azepan-2-one

(E)-3-(2-chlorobenzylidene)azepan-2-one

Made according to literature. TFAA was added to a solution ofazepan-2-one (0.5 g, 4.4 mmol) in toluene at 0° C., after stirring underice bath for 1 h, the solvent was removed to give a residue. The residuewas mixed with 2-chlorobenzaldehyde (0.558 g, 3.98 mmol) and charged toa suspension of t-BuOK (0.596 mg, 5.3 mmol) in THF at 0° C., theresulting mixture was then heated to 50-60° C. and stirred at thistemperature for 1.5 h. The reaction mixture was concentrated undervacuum and water was then added. The resulting mixture was extractedwith EA for 3 times, the combined organic phase was washed with brineand dried over anhydrous sodium sulfate. Evaporation of solvent gave aresidue which was purified through column to get product (0.6 g, 60%yield).

¹H NMR (400 MHz, CDCl₃): δ 7.407-7.431 (m, 1H), 7.314-7.338 (m, 1H),7.173-7.280 (m, 2H), 7.173 (s, 1H), 6.392 (br, 1H), 3.299 (m, 2H), 2.423(m, 2H), 1.761 (m, 4H); ¹³CNMR (100 MHz, CDCl₃) δ 175.773, 140.278,134.708, 134.116, 131.748, 130.109, 129.714, 129.062, 126.595, 42.463,28.764, 27.687, 27.118.

(E)-3-(2-chlorobenzylidene)-1-((E)-3-(3,4,5-trimethoxyphenyl)acryloyl)azepan-2-one

To a solution of (E)-3-(2-chlorobenzylidene)azepan-2-one (24 mg, 0.102mmol) in THF at −78° C. was added 1.6 M n-BuLi (70 ul, 0.112 mmol),after stirring at −78° C. for 1 h, (E)-tert-butyl3-(3,4,5-trimethoxyphenyl)acrylate (30 mg, 0.103 mmol) was added and theresulting mixture was stirred at −78° C. for additional 1 h. Thereaction was quenched with saturated NH₄Cl solution and then extractedwith EA for 3 times, the combined organic phase was washed with brineand dried over anhydrous sodium sulfate. Evaporation of solvent gave aresidue which was purified through column to get pure product as whitesolid (30 mg, 65% yield).

¹H NMR (400 MHz, CDCl₃): δ 7.735 (d, J=15.2 Hz, 1H), 7.449-7.473 (m,1H), 7.375-7.411 (m, 3H), 7.293-7.316 (m, 2H), 6.819 (s, 2H), 4.020 (m,2H), 3.918 (s, 6H), 3.888 (s, 3H), 2.535 (m, 2H), 1.809 (m, 4H); ¹³CNMR(100 MHz, CDCl₃) δ 175.097, 167.759, 153.521, 144.536, 140.954, 140.142,134.587, 134.260, 133.949, 130.747, 129.965, 129.790, 129.722, 126.823,119.909, 105.634, 61.117, 56.373, 44.018, 27.831, 27.649, 27.277.

REFERENCES

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What is claimed is:
 1. A method of selectively killing one or moresenescent cells from a cell population or tissue, the method comprisingcontacting the senescent cells with an effective amount of a compositioncomprising a pharmaceutically acceptable carrier and a compoundcomprising Formula (II):

wherein: X is selected from the group consisting of C(O), C(S), C(NH)and S(O)₂; Y is selected from the group consisting of O, NH and S; n is1; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are each independently selectedfrom the group consisting of hydrogen, deuterium, halogen, CF₃, CN, OH,OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR, NO₂, CONRR,OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted C1 to C6 alkenyl, a substituted or unsubstituted C1 to C6alkynyl, and a substituted or unsubstituted aryl; R is independentlyselected from the group consisting of hydrogen, substituted C1-C4aliphatic moiety, aliphatic moiety containing nitrogen, oxygen, orsulfur, or alternately, two R moieties bound to the same nitrogen atomare optionally taken together with the nitrogen atom to form a 3-7membered saturated or unsaturated ring having 1-2 additional heteroatomsindependently selected from the group consisting of nitrogen, oxygen, orsulfur; A is selected from the group consisting of

wherein: R₉ and R₁₀ are independently selected from the group consistingof hydrogen, deuterium, halogen, CF₃, CN, OH, OCH₃, OR′, SR′, NR′R′,NR′COR′, NR′CONR′R′, NR′CO₂R′, COR′, CO₂R′, NOR′, NO₂, CONR′R′,OC(O)NR′R′, SO₂R, SO₂NR′R′, NR′SO₂R′, NR′SO₂NR′R′, C(O)C(O)R′, andC(O)CH₂C(O)R′, a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted C1 to C6 alkenyl, a substituted orunsubstituted C1 to C6 alkynyl, and a substituted or unsubstituted aryl;R′ is independently selected from the group consisting of hydrogen,substituted C1-C4 aliphatic moiety, aliphatic moiety containingnitrogen, oxygen, or sulfur, or alternately, two R′ moieties bound tothe same nitrogen atom are optionally taken together with the nitrogenatom to form a 3-7 membered saturated or unsaturated ring having 1-2additional heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; Optionally, R₉ and R₁₀ are takentogether to form a 5-8 membered saturated or unsaturated ring having 0-3heteroatoms independently selected from the group consisting ofnitrogen, oxygen, or sulfur; B is

wherein: R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅ are each independently selectedfrom the group consisting of hydrogen, deuterium, halogen, CF₃, CN, OH,OCH₃, OR″, SR″, NR″R″, NR″COR″, NR″CONR″R″, NRCO₂R″, COR″, CO₂R″, NOR″,NO₂, CONR″R″, OC(O)NR″R″, SO₂R″, SO₂NR″R″, NR″SO₂R″, NR″SO₂NR″R″,C(O)C(O)R″, and C(O)CH₂C(O)R″, a substituted or unsubstituted C1 to C6alkyl, a substituted or unsubstituted C1 to C6 alkenyl, a substituted orunsubstituted C1 to C6 alkynyl, and a substituted or unsubstituted aryl;R″ is independently selected from the group consisting of hydrogen,substituted C1-C4 aliphatic moiety, aliphatic moiety containingnitrogen, oxygen, or sulfur, or alternately, two R″ moieties bound tothe same nitrogen atom are optionally taken together with the nitrogenatom to form a 3-7 membered saturated or unsaturated ring having 1-2additional heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; Optionally, R₁₁ and R₁₂, R₁₂ and R₁₃,R₁₃ and R₁₄, and R₁₄ and R₁₅ are taken together to form a 4-8 memberedsaturated or unsaturated ring having 0-3 heteroatoms independentlyselected from the group consisting of nitrogen, oxygen, or sulfur; m isan integer from 0-6; and or B is one or more monocyclic aryl, a 3-7membered saturated or partially unsaturated carbocyclic ring, an 8-10membered bicyclic saturated, partially unsaturated or aryl ring, a 5-6membered monocyclic heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, a 4-7 membered saturated orpartially unsaturated heterocyclic ring having 1-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, a 7-10 memberedbicyclic saturated or partially unsaturated heterocyclic ring having 1-5heteroatoms independently selected from the group consisting ofnitrogen, oxygen or sulfur, or an 8-10 membered bicyclic heteroaryl ringhaving 1-5 heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; wherein any and all of the substituteddescribed are substituted with one or more groups that are independentlya methyl group or methoxy group.
 2. The method of claim 1, wherein thesenescent cells are senescent due to replicative cellular senescence,premature cellular senescence, or therapy-induced senescence.
 3. Themethod of claim 1, wherein the senescent cells are from an age-relatedpathology.
 4. The method of claim 1, wherein the composition furthercomprises at least one inhibitor of one or more anti-apoptotic proteinsin the Bcl-2 family.
 5. The method of claim 4, wherein the inhibitor isABT263.
 6. The method of claim 1, wherein the cell population or tissueis in a human subject.
 7. The method of claim 6, wherein the subject hasa received DNA-damaging therapy.
 8. A method of killing therapy-inducedsenescent cells from a cell population or tissue, the method comprisingcontacting the therapy-induced senescent cells with an effective amountof a composition comprising a pharmaceutically acceptable carrier and acompound comprising formula (II):

wherein: X is selected from the group consisting of C(O), C(S), C(NH)and S(O)₂; Y is selected from the group consisting of O, NH and S; n is1; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are each independently selectedfrom the group consisting of hydrogen, deuterium, halogen, CF₃, CN, OH,OCH₃, OR, SR, NRR, NRCOR, NRCONRR, NRCO₂R, COR, CO₂R, NOR, NO₂, CONRR,OC(O)NRR, SO₂R, SO₂NRR, NRSO₂R, NRSO₂NRR, C(O)C(O)R, and C(O)CH₂C(O)R, asubstituted or unsubstituted C1 to C6 alkyl, a substituted orunsubstituted C1 to C6 alkenyl, a substituted or unsubstituted C1 to C6alkynyl, and a substituted or unsubstituted aryl; R is independentlyselected from the group consisting of hydrogen, substituted C1-C4aliphatic moiety, aliphatic moiety containing nitrogen, oxygen, orsulfur, or alternately, two R moieties bound to the same nitrogen atomare optionally taken together with the nitrogen atom to form a 3-7membered saturated or unsaturated ring having 1-2 additional heteroatomsindependently selected from the group consisting of nitrogen, oxygen, orsulfur; A is selected from the group consisting of

wherein: R₉ and R₁₀ are independently selected from the group consistingof hydrogen, deuterium, halogen, CF₃, CN, OH, OCH₃, OR′, SR′, NR′R′,NR′COR′, NR′CONR′R′, NR′CO₂R′, COR′, CO₂R′, NOR′, NO₂, CONR′R′,OC(O)NR′R′, SO₂R, SO₂NR′R′, NR′SO₂R′, NR′SO₂NR′R′, C(O)C(O)R′, andC(O)CH₂C(O)R′, a substituted or unsubstituted C1 to C6 alkyl, asubstituted or unsubstituted C1 to C6 alkenyl, a substituted orunsubstituted C1 to C6 alkynyl, and a substituted or unsubstituted aryl;R′ is independently selected from the group consisting of hydrogen,substituted C1-C4 aliphatic moiety, aliphatic moiety containingnitrogen, oxygen, or sulfur, or alternately, two R′ moieties bound tothe same nitrogen atom are optionally taken together with the nitrogenatom to form a 3-7 membered saturated or unsaturated ring having 1-2additional heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; Optionally, R₉ and R₁₀ are takentogether to form a 5-8 membered saturated or unsaturated ring having 0-3heteroatoms independently selected from the group consisting ofnitrogen, oxygen, or sulfur; B is

wherein: R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅ are each independently selectedfrom the group consisting of hydrogen, deuterium, halogen, CF₃, CN, OH,OCH₃, OR″, SR″, NR″R″, NR″COR″, NR″CONR″R″, NRCO₂R″, COR″, CO₂R″, NOR″,NO₂, CONR″R″, OC(O)NR″R″, SO₂R″, SO₂NR″R″, NR″SO₂R″, NR″SO₂NR″R″,C(O)C(O)R″, and C(O)CH₂C(O)R″, a substituted or unsubstituted C1 to C6alkyl, a substituted or unsubstituted C1 to C6 alkenyl, a substituted orunsubstituted C1 to C6 alkynyl, and a substituted or unsubstituted aryl;R″ is independently selected from the group consisting of hydrogen,substituted C1-C4 aliphatic moiety, aliphatic moiety containingnitrogen, oxygen, or sulfur, or alternately, two R″ moieties bound tothe same nitrogen atom are optionally taken together with the nitrogenatom to form a 3-7 membered saturated or unsaturated ring having 1-2additional heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; Optionally, R₁₁ and R₁₂, R₁₂ and R₁₃,R₁₃ and R₁₄, and R₁₄ and R₁₅ are taken together to form a 4-8 memberedsaturated or unsaturated ring having 0-3 heteroatoms independentlyselected from the group consisting of nitrogen, oxygen, or sulfur; m isan integer from 0-6; and or B is one or more monocyclic aryl, a 3-7membered saturated or partially unsaturated carbocyclic ring, an 8-10membered bicyclic saturated, partially unsaturated or aryl ring, a 5-6membered monocyclic heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, a 4-7 membered saturated orpartially unsaturated heterocyclic ring having 1-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, a 7-10 memberedbicyclic saturated or partially unsaturated heterocyclic ring having 1-5heteroatoms independently selected from the group consisting ofnitrogen, oxygen or sulfur, or an 8-10 membered bicyclic heteroaryl ringhaving 1-5 heteroatoms independently selected from the group consistingof nitrogen, oxygen, or sulfur; wherein any and all of the substituteddescribed are substituted with one or more groups that are independentlya methyl group or methoxy group.
 9. The method of claim 8, wherein thecomposition further comprises at least one inhibitor of one or moreanti-apoptotic proteins in the Bcl-2 family.
 10. The method of claim 9,wherein the inhibitor is ABT263.
 11. The method of claim 8, wherein thetherapy-induced senescent cells are in a human subject.